This week the Node is in Boston, MA! This Friday (25th of September) our community manager Cat will be giving two talks:

2.30 p.m.- Inside a Career in Scientific Publishing and Science Communication

Tufts Boston Campus, Sackler Building (145 Harrison Avenue), Room 316

6 p.m.- YEN Boston discussion: Building a Developmental Biology Community

Harvard Medical School, Warren Alpert Building: Room 563 (register here)

If you are in the Boston area, join us for one (or both!) of these talks! Cat would love to chat to you before or after the talks about the Node, social media or alternative careers. We look forward to meeting you!

(2 votes)

Loading...

This editorial first appeared in Development.

In September 2014, Development organised a four-day workshop titled ‘From Stem Cells to Human Development’. In planning this meeting, we sought to fill what we saw as a gap in the meeting calendar – a way of bringing together a diverse cross-section of researchers with a common interest in using the rapidly developing tools of stem cell biology, genetic engineering and genomic analysis to understand human development (for a review of the meeting, see Medvinsky and Livesey, 2015). The enthusiasm with which this workshop was met, from invited speakers and registered participants alike, confirmed our view that we are now in a period in which significant inroads into understanding human development will be made. With our ever-improving ability to model tissue development in vitro and to manipulate the human genome (and epigenome), we are now in a position to analyse human organogenesis and to understand how it differs from that in other model organisms – and hence to start to probe the developmental biology underlying the evolution of our species.

As we wrote in an editorial in January, “The human development field represents an essential growth area for the developmental biology community, and Development is keen to play an active role in supporting and inspiring it” (Pourquié et al., 2015). This Special Issue celebrates that aim – bringing together a collection of Reviews and Research Articles that directly address a broad range of topics in human developmental biology: from the earliest stages of human development to cellular ageing and degeneration, and from basic questions of how an organ is formed to ways in which we might translate this knowledge in the clinic. We are also supporting this initiative with a second ‘From Stem Cells to Human Development’ meeting, to be held in September 2016. More details on what should be a fantastic follow-up event can be found at http://workshops.biologists.com/from-stem-cells-to-human-development-2/.

Studying human development is obviously a challenging endeavour, given the practical and ethical difficulties in working with human material. However, as discussed by Dianne Gerrelli and colleagues (2015), there is a growing set of resources for researchers, including the Human Developmental Biology Resource (with which the authors are affiliated), which provides embryonic and foetal material and a range of valuable services. Maintaining and developing such resources will be essential as research on human development progresses.

Complementing work using human tissue, much of the research into human development relies on the generation and manipulation of human pluripotent stem cells (hPSCs) – either embryonic (hESCs) or induced (hiPSCs). There has, however, been much debate surrounding the pluripotent status of such hPSCs, particularly when compared with their mouse equivalents, as well as their in vivo counterparts. In their Review, Martin Pera and colleagues (Davidson et al., 2015) discuss these controversies in the light of recent attempts to generate truly naïve hESCs. Kathy Niakan and co-workers are also interested in pluripotent states and human-mouse comparisons. In their Research Article (Blakeley et al., 2015), they report single-cell RNA sequencing analyses of human and mouse preimplantation epiblasts, identifying important differences in the transcriptomes – and presumably therefore the development – of the early human and mouse embryo. One challenge in the field has been that functional assays for pluripotency of human cells are limited. To address this, Hiromitsu Nakauchi and colleagues (Masaki et al., 2015) investigate whether generating inter-specific chimeras (using mouse epiblasts and PSCs from various species) might provide an alternative assay system. Also using mouse embryology to probe human development are Felipe Vilella and colleagues, who describe a microRNA secreted in human endometrial fluid that can promote mouse embryo adhesion during implantation (Vilella et al., 2015), potentially identifying a novel route by which the efficiency of implantation can be modulated.

In another research paper investigating the role of microRNAs in human development (Jönsson et al., 2015), Malin Parmar and co-workers analyse the microRNAs expressed in the human foetal brain and in PSC-derived neural progenitor cells, identifying region-specific microRNAs that probably influence neural cell fate. Generating a functional nervous system requires not only that cell fate is correctly defined, but also that appropriate connectivity is established and that neurons are properly supported by glia. Frederick Livesey and colleagues address the former problem in cortical neuron cultures (Kirwan et al., 2015), while Motoharu Sakaue and Maya Sieber-Blum describe a protocol for generating supporting Schwann-like cells from human epidermal neural crest stem cells (Sakaue and Sieber-Blum, 2015). Meanwhile, Ikuo Suzuki and Pierre Vanderhaeghen (2015) review various aspects of studying neural development using hPSCs and discuss how these approaches should allow us to gain insights into the evolution of the human brain.

Katie Pollard and Lucia Franchini’s interests also lie in understanding human evolution, but from a genomic perspective. Their Review (Franchini and Pollard, 2015) discusses how we can combine sequencing information with functional genomics and stem cell biology to identify and characterise changes in the human genome that might have led to human-specific developmental traits. They highlight the importance of appropriate experimental systems – not only model organisms but also through human stem cells and organoids – in which to test the function of human-specific genomic features. The ability to model not only cellular differentiation but also tissue formation in a dish constitutes a major breakthrough in the field over the past decade. Meritxell Huch and Bon-Kyoung Koo review the latest advances in generating endodermal organoids from both embryonic and adult stem cells (Huch and Koo, 2015) and provide a perspective on where this field is heading. The Review by Neil Hanley and colleagues (Jennings et al., 2015), while also focussing on endoderm development – in this case, pancreas – provides a complementary viewpoint, discussing what is known about human pancreas development in vivo and how these insights translate into our ability to generate β-cells in vitro.

Turning to other organs, Christine Mummery and Charles Murry and their colleagues both focus on heart development. Mummery’s work (van den Berg et al., 2015) characterises the transcriptome of the human foetal heart and compares it with the RNA profile of PSC-derived cardiomyocytes. Meanwhile, Murry’s study (Palpant et al., 2015) provides insights into how cardiomyocytes are specified in a hESC system. The final Research Article of this issue returns to the topic of organoid formation, this time the mammary gland. Christina Scheel and co-workers (Linnemann et al., 2015) present an organoid system that allows the regenerative potential and morphogenetic dynamics of mammary epithelial cells to be studied.

Although understanding human development is an important goal in itself, the translational potential of this field is clear: if we can grow human tissues in vitro, we can use these to model disease, to test potential drugs and to develop cell therapies. Two Spotlights in this issue discuss these aspects of the field. Scott Thies and Charles Murry (2015) present some of the most promising preclinical data and clinical trials of stem cell therapies, while Elsa Vera and Lorenz Studer (2015) highlight a potential problem with using stem cell-derived models in disease research: both hESCs and hiPSCs are ‘young’ cells, whereas many diseases – particularly neurodegenerative disorders – afflict the old. Although these articles stray from the classic scope of a developmental biology journal, we hope that they illustrate the continuum of both the field, from basic understanding of developmental processes to their applications in regenerative therapy, and of development itself – from embryogenesis through post-embryonic maturation to ageing and decline.

We have a limited number of print copies of this Special Issue to give away to interested readers. If you would like one, please send an email to dev@biologists.com with your mailing address. Whether in print or online, we hope you enjoy this Special Issue on Human Development. We see an exciting future for this field, and we want Development to be at the heart of it. We therefore encourage those of you working in this area to consider Development as a potential venue for the publication of your best work and we look forward to many more exciting human development papers finding their way into the pages of our journal.

(3 votes)

Loading...

Here are the highlights from the current issue of Development – our Special Issue on “Human Development”. This Special Issue brings together a collection of Reviews and Research Articles that directly address a broad range of topics in human developmental biology: from the earliest stages of human development to cellular ageing and degeneration, and from basic questions of how an organ is formed to ways in which we might translate this knowledge in the clinic. Happy reading!

Enabling research with human embryonic and fetal tissue resources

Gerrelli and colleagues summarise the remit and efforts of the Human Developmental Biology Resource – and other similar projects – that provides researchers with access to human tissue and related services. See the Spotlight article on p. 3073

The advancement of human pluripotent stem cell-derived therapies into the clinic

In their opinion piece, Thies and Murry consider progress in the use of human PSCs in regenerative medicine – fuelled by advances in developmental biology – in five areas that offer great promise for clinical applications. See the Spotlight article on p. 3077

When rejuvenation is a problem: challenges of modeling late-onset neurodegenerative disease

Vera and Studer discuss the difficulties involved in using stem cells to study diseases of the elderly and review how these might be overcome. See the Spotlight article on p. 3085

The pluripotent state in mouse and human

Pera and colleagues examine recent efforts in defining and capturing the human naïve pluripotent state in vivo and in vitro, in light of the different states of pluripotency found in the mouse. See the Review article on p. 3090

Genomic approaches to studying human-specific developmental traits

Franchini and Pollard discuss how genome sequencing and functional genomic approaches are enabling analyses of the evolutionary and developmental origin of traits unique to our species. See the Review article on p. 3100

Modelling mouse and human development using organoid cultures

Huch and Koo review recent advances in the generation of ESC- and adult stem cell-derived organoids in order to understand the development of endoderm-derived organs in human and to develop therapeutic strategies for repair. See the Review article on p. 3113

Human pancreas development

Jennings, Hanley and colleagues present a human-centric view of the latest advances in our understanding of pancreas development and the relevance of these insights from a clinical perspective. See the Review article on p. 3126

Is this a brain which I see before me? Modeling human neural development with pluripotent stem cells

Suzuki and Vanderhaeghen examine stem cell-based approaches to analysing human brain development, from specification of particular cell types to building neuronal networks. See the Review article on p. 3138

PLUS:

Research Articles/Techniques and Resources:

• Defining the three cell lineages of the human blastocyst by single-cell RNA-seq

• Comprehensive analysis of microRNA expression in regionalized human neural progenitor cells reveals microRNA-10 as a caudalizing factor

• Development and function of human cerebral cortex neural networks from pluripotent stem cells in vitro

• Human epidermal neural crest stem cells as a source of Schwann cells

• Inhibition of β-catenin signaling respecifies anterior-like endothelium into beating human cardiomyocytes

• Hsa-miR-30d, secreted by the human endometrium, is taken up by the pre-implantation embryo and might modify its transcriptome

• Interspecific in vitro assay for the chimera-forming ability of human pluripotent stem cells

• Transcriptome of human foetal heart compared with cardiomyocytes from pluripotent stem cells

• Quantification of regenerative potential in primary human mammary epithelial cells

(No Ratings Yet)

Loading...

The National University of Singapore (NUS) invites applications for faculty appointment as Head of the Department of Anatomy.

The Department’s mission is to enhance the international stature of the School and of NUS through excellence in teaching and research. It is committed to teaching undergraduate medical, dental, pharmacy, and life science students. In addition, the Department provides research training to both undergraduate and postgraduate research students. Currently, research in the Department focuses on neurobiology, cancer biology and venoms & toxins in a vibrant academic environment. Researchers in the Department use a variety of techniques in cellular and molecular biology and the Department and the School are well equipped with state-of-the-art facilities.

The Department has over the past 5 years attracted SGD 10 million in competitive grant funding. The Department has 21 faculty members and a total of around 100 staff and postgraduate students. The Department publishes on average 57 scientific papers annually in high impact journals such as Cell, Nature Cell Biology, Nature Medicine, Journal of Clinical Investigations, Cancer Research, Hepatology, Advanced Materials, PNAS and Journal of Neuroscience. The Department enjoys strong collaborative links with research institutes within NUS, government agencies as well as overseas research institutes and leading universities.

The candidate should be an outstanding scholar who will be able to provide strong leadership in research and teaching with an excellent track record and international recognition for research in any of the main areas of research in the Department, or in developmental biology. Administrative experience in leading an academic department would be an added advantage. A generous start up package and first class laboratory facilities are available. The Head is expected to generate strong research programmes, secure external funding and provide intellectual leadership characteristic of a world-class university.

Remuneration will commensurate with the candidate’s qualifications and experience. Informal enquiries can be made to Ms Lee Sing Ee: medlse@nus.edu.sg Tel +65 6772 3729.

Interested parties should submit their applications, supported by a detailed resume and names of at least six referees to:

Ms LEE Sing Ee

Assistant Director, Academic Affairs

Dean’s Office

Yong Loo Lin School of Medicine

National University of Singapore

1E Kent Ridge Road, NUHS Tower Block

Level 11, Singapore 119228

Fax: +65-6778-5743   Email: medlse@nus.edu.sg

Closing Date: 31 October 2015

(Only shortlisted candidates will be notified)

(No Ratings Yet)

Loading...

Genetically engineered mouse models have been used extensively to study a wide variety of biological processes in vivo, and innovations in genetic engineering have made it possible to dissect more intricate biological questions. For example, the first mice that showed successful germline transmission of foreign DNA were created in the 1980s, and this allowed the generation of various transgenic “oncomice” such as the first mouse model of breast cancer using the MMTV-Myc transgene ¹, and the first mouse model of osteosarcoma using the MT-FosLTR transgene ². Subsequently in the early 1990s, mice were developed to have whole-body deletions of various genes such as the p53 tumor suppressor gene ³. The combination of the ability to introduce oncogenes and delete tumor suppressor genes through these initial transgenic mouse technologies contributed greatly to our current understanding of cancer development, progression, and treatment.

Nonetheless, transgenic mice and whole-body knock out mice sometimes do not survive development ⁴. To overcome this limitation, investigators harnessed site-specific recombinase systems from bacteriophages and yeasts such as the Cre-loxP and the Flp-FRT systems, which were first adapted to the mouse genome for germline transmission in the 1990s to enable temporally-regulated and tissue-specific genetic manipulations ^{5, 6}. Since then, many conditional mouse alleles utilizing transgenic or endogenous promoter-driven Cre and loxP-regulated genes have been generated to increase tissue-specificity of gene expression and decrease pre-mature lethality and other unwanted phenotypes. Additionally, replication-defective viruses (adenoviruses, lentiviruses) containing Cre recombinase (Adeno-Cre, Lenti-Cre) have also been used to further improve upon the temporal regulation of gene expression ⁷. Further modifications enabled the viruses to carry specific promoter-driven Cre recombinases to add tissue specificity after viral delivery ⁸. Finally, similar temporal regulation can be achieved by using fusion proteins combining Cre and mutated hormone receptors such as the estrogen receptor (Cre-ER^T2) ⁹. By utilizing this approach, metabolites of tamoxifen can be used to translocate Cre-ER^T2 to the nucleus and thus activate subsequent Cre-mediated gene modification.

Recently, there has been increasing interest in combining more than one recombinase system in the same mouse model using dual recombinase technology to allow for sequential mutagenesis and to better model sporadic cancers in the adult mouse ^{10, 11}. In our lab, we have generated two complimentary new mouse alleles that facilitates the regulation of Cre-ER^T2 by the Flp-FRT recombinase system. Both alleles are knocked into the endogenous Col1a1 locus for ubiquitous expression, and also at the same time reserve the two Rosa26 alleles for other transgenes of interest. In the first mouse allele, Col1a1^{FRT-Cre-ER-T2-FRT}, Cre-ER^T2 is flanked by FRT sites. The rationale to generate Col1a1^{FRT-Cre-ER-T2-FRT} mice is to enable whole animal ubiquitous expression of Cre-ER^T2 until exposure to Flp recombinase (Figure 1A). After Flp-mediated recombination of the FRT sites, cells are no longer able to express Cre-ER^T2 and therefore lose the ability to delete DNA flanked by loxP sites following exposure to tamoxifen. In this way, different mutations can be introduced in adjacent cells in vivo so that the consequences for intercellular interactions, such as cancer cells and stromal cells, can be studied.   In the second mouse allele, Col1a1^{FRT-STOP-FRT-Cre-ER-T2}, Cre-ER^T2 sits downstream of a FRT-flanked STOP cassette, which inhibits transcription of Cre-ER^T2. The rationale to generate Col1a1^{FRT-STOP-FRT-Cre-ER-T2} is that initially no cell expresses Cre-ER^T2 because transcription of Cre-ER^T2 is terminated by an upstream FRT site-flanked transcription STOP cassette (Figure 1B). However, after Flp-mediated recombination, the STOP cassette is excised. Therefore, these cells can initiate transcription of the Cre-ER^T2 fusion protein, which in response to subsequent exposure to tamoxifen translocates into the nucleus to recombine DNA flanked by loxP sites. Cells without exposure to Flp will not be able to undergo Cre-mediated DNA recombination. In this way, the Col1a1^{FRT-STOP-FRT-Cre-ER-T2} allele enables sequential mutations within the same cell over time. First, one mutation occurs in the cell from Flp recombinase, and then tamoxifen activates Cre recombinase in the same cell to mutate a second gene to study how the order of gene mutations may affect cellular outcome. In addition, multiple genes may be mutated by Flp recombinase to initiate tumor development. Then, the role of a loxP-flanked gene in tumor maintenance can be studied because only the tumor cell will express Cre-ER^T2. This allele can therefore be used to identify potential therapeutic targets.

Through characterization of the Col1a1^{FRT-Cre-ER-T2-FRT} mice using the reporter allele Rosa26^{mTmG 12}, we found that a single intraperitoneal injection of tamoxifen at 75mg/kg into Col1a1^{FRT-Cre-ER-T2-FRT}; Rosa26^mTmG/+ mice is sufficient to translocate the Cre-ER^T2 fusion protein into the nucleus and mediate recombination of the loxP-sites flanking tdTomato. This resulted in the deletion of the tdTomato red fluorescent protein, and the subsequent expression of eGFP green fluorescent protein from cells of all tissues examined. The limitation of this model is that there is an age- and tissue-dependent Cre-ER^T2 activation independent of tamoxifen administration, most notably in the pancreas and the liver. Therefore, in experiments involving these organs using the Col1a1^{FRT-Cre-ER-T2-FRT} mice, age of the mice at the onset of Flp and tamoxifen administration is crucial for the interpretation of the results.

We also characterized the Col1a1^{FRT-STOP-FRT-Cre-ER-T2} mice using the reporter allele Rosa26^mTmG. When Col1a1^{FRT-STOP-FRT-Cre-ER-T2}; Rosa26^mTmG/+ mice were given a single intraperitoneal injection of tamoxifen at 75mg/kg, there was no Cre-ER^T2-mediated deletion of the tdTomato red fluorescent protein or expression of the eGFP green fluorescent protein. Next, we generated a mouse model of soft tissue sarcoma using the new allele to verify the ability of the STOP cassette to be removed by Flp, and the subsequent ability of Cre-ER^T2 to manipulate loxP-flanked alleles. In this model, Col1a1^{FRT-STOP-FRT-Cre-ER-T2}; Kras^{FRT-STOP-FRT/+}; p53^FRT/FRT; Rosa26^mTmG/+ mice were first administered intramuscular injection of adenovirus carrying Flp recombinase in the hindlimb to form soft tissue sarcomas at the site of injection via the expression of the oncogenic KRAS protein and elimination of the p53 tumor suppressor protein ^{11, 13, 14}. Following the formation of soft tissue sarcomas, either single intratumoral injection of 4-hydroxytamoxifen or several doses of systemic 4-hydroxytamoxifen were administered. The resulting tumor showed tumor parenchymal cells with deletion of tdTomato red fluorescent protein and expression of eGFP green fluorescent protein, while the tumor stromal cells such as the vasculature continued to express tdTomato. One potential challenge in this model is that while the Col1a1^{FRT-STOP-FRT-Cre-ER-T2} is functional, single intraperitoneal injection of tamoxifen failed to activate Cre-ER^T2 in primary sarcomas. Therefore, the delivery of an adequate level of tamoxifen and/or its metabolites into the tumor is critical to activate Cre-ER^T2 expression with this allele.

We anticipate that these two new alleles will allow for increased control of gene manipulation in genetically engineered mouse models. These alleles, in conjunction with other new technologies in the field such as the CRISPR/Cas9 system ¹⁵, have the potential to bring together the speed and efficiency of the CRISPR/Cas9 system with the spatial and temporal control of dual recombinase technology, manipulate the genome in vivo to study development, cancer and other diseases in the mouse.

Minsi Zhang and David Kirsch, Department of Radiation Oncology, Duke University Medical Center

Main reference

Zhang, M., & Kirsch, D. The generation and characterization of novel Col1a1FRT-Cre-ER-T2-FRT and Col1a1FRT-STOP-FRT-Cre-ER-T2 mice for sequential mutagenesis. Disease Models & Mechanisms. 2015. 8(9), 1155-1166 DOI: 10.1242/dmm.021204

Other references:

1. Stewart TA, Pattengale PK, Leder P. Spontaneous mammary adenocarcinomas in transgenic mice that carry and express MTV/myc fusion genes. Cell. 1984;38(3):627-37. PubMed PMID: 6488314.
2. Ruther U, Komitowski D, Schubert FR, Wagner EF. c-fos expression induces bone tumors in transgenic mice. Oncogene. 1989;4(7):861-5. PubMed PMID: 2547184.
3. Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery CA, Jr., Butel JS, Bradley A. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 1992;356(6366):215-21. doi: 10.1038/356215a0. PubMed PMID: 1552940.
4. Jacks T, Fazeli A, Schmitt EM, Bronson RT, Goodell MA, Weinberg RA. Effects of an Rb mutation in the mouse. Nature. 1992;359(6393):295-300. doi: 10.1038/359295a0. PubMed PMID: 1406933.
5. Orban PC, Chui D, Marth JD. Tissue- and site-specific DNA recombination in transgenic mice. Proc Natl Acad Sci U S A. 1992;89(15):6861-5. PubMed PMID: 1495975; PubMed Central PMCID: PMC49604.
6. Dymecki SM. Flp recombinase promotes site-specific DNA recombination in embryonic stem cells and transgenic mice. Proc Natl Acad Sci U S A. 1996;93(12):6191-6. PubMed PMID: 8650242; PubMed Central PMCID: PMC39212.
7. Wang Y, Krushel LA, Edelman GM. Targeted DNA recombination in vivo using an adenovirus carrying the cre recombinase gene. Proc Natl Acad Sci U S A. 1996;93(9):3932-6. PubMed PMID: 8632992; PubMed Central PMCID: PMC39462.
8. Sutherland KD, Proost N, Brouns I, Adriaensen D, Song JY, Berns A. Cell of origin of small cell lung cancer: inactivation of Trp53 and Rb1 in distinct cell types of adult mouse lung. Cancer Cell. 2011;19(6):754-64. doi: 10.1016/j.ccr.2011.04.019. PubMed PMID: 21665149.
9. Indra AK, Warot X, Brocard J, Bornert JM, Xiao JH, Chambon P, Metzger D. Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases. Nucleic Acids Res. 1999;27(22):4324-7. PubMed PMID: 10536138; PubMed Central PMCID: PMCPMC148712.
10. Shai A, Dankort D, Juan J, Green S, McMahon M. TP53 Silencing Bypasses Growth Arrest of BRAFV600E-Induced Lung Tumor Cells In a Two-Switch Model of Lung Tumorigenesis. Cancer research. 2015. doi: 10.1158/0008-5472.CAN-14-3701. PubMed PMID: 26001956.
11. Schonhuber N, Seidler B, Schuck K, Veltkamp C, Schachtler C, Zukowska M, Eser S, Feyerabend TB, Paul MC, Eser P, Klein S, Lowy AM, Banerjee R, Yang F, Lee CL, Moding EJ, Kirsch DG, Scheideler A, Alessi DR, Varela I, Bradley A, Kind A, Schnieke AE, Rodewald HR, Rad R, Schmid RM, Schneider G, Saur D. A next-generation dual-recombinase system for time- and host-specific targeting of pancreatic cancer. Nat Med. 2014;20(11):1340-7. doi: 10.1038/nm.3646. PubMed PMID: 25326799; PubMed Central PMCID: PMC4270133.
12. Muzumdar MD, Tasic B, Miyamichi K, Li L, Luo L. A global double-fluorescent Cre reporter mouse. Genesis. 2007;45(9):593-605. doi: 10.1002/dvg.20335. PubMed PMID: 17868096.
13. Moding EJ, Lee CL, Castle KD, Oh P, Mao L, Zha S, Min HD, Ma Y, Das S, Kirsch DG. Atm deletion with dual recombinase technology preferentially radiosensitizes tumor endothelium. J Clin Invest. 2014;124(8):3325-38. doi: 10.1172/JCI73932. PubMed PMID: 25036710; PubMed Central PMCID: PMC4109553.
14. Moding EJ, Castle KD, Perez BA, Oh P, Min HD, Norris H, Ma Y, Cardona DM, Lee CL, Kirsch DG. Tumor cells, but not endothelial cells, mediate eradication of primary sarcomas by stereotactic body radiation therapy. Sci Transl Med. 2015;7(278):278ra34. doi: 10.1126/scitranslmed.aaa4214. PubMed PMID: 25761890; PubMed Central PMCID: PMC4360135.
15. Platt RJ, Chen S, Zhou Y, Yim MJ, Swiech L, Kempton HR, Dahlman JE, Parnas O, Eisenhaure TM, Jovanovic M, Graham DB, Jhunjhunwala S, Heidenreich M, Xavier RJ, Langer R, Anderson DG, Hacohen N, Regev A, Feng G, Sharp PA, Zhang F. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell. 2014;159(2):440-55. doi: 10.1016/j.cell.2014.09.014. PubMed PMID: 25263330; PubMed Central PMCID: PMC4265475.

(1 votes)

Loading...

PhD POSITION

THE ROLE OF JNK SIGNALLING IN BRAIN REGENERATION

A PhD position is currently available (starting January 2016) to develop a project involving the analysis of the role(s) of the JNK signalling in brain regeneration using Drosophila (mainly) and zebrafish as model systems.

Most of the project will be developed at the “Cell Signalling and Morphogenesis” laboratory of the Institute of Molecular Biology of Barcelona but will also include short periods at the “Center for Regenerative Therapies TU Dresden”

REQUIREMENTS

Graduates in Biology, Physics or related areas with a strong track record and deep interest in Developmental Genetics, Neurosciences or Regenerative Biomedicine are encouraged to apply. Laboratory experience would be a definitive advantage.

The successful candidate will hold a research stipend in Dresden for up to 3 years and develop his/her research project(s) using a wide array of techniques and in vivo models.

CONTACT:

Dr. Enrique Martin‐Blanco, IBMB‐CSIC, SPAIN

(+34934034668 – embbmc [at] ibmb.csic.es)

Prof. Michael Brand, CRTD, GERMANY

(+4935145882301 – michael.brand [at] biotec.tu-dresden.de)

Job offer EMB-MB

(No Ratings Yet)

Loading...

This Sticky Wicket article first featured in Journal of Cell Science. Read other articles and cartoons of Mole & Friends here.

Text translated from the Italian. I think. With profound apologies to Dante Alighieri (D.A.)

Midway (or more? I hope not) on the journey of my independent career

I found myself in a dark forest…okay, it was my dark office;

My grant had been rejected, and the revision was soon due.¹

I wasn’t writing normally; everything was coming out in triplets.

Sort of like a poem, except it didn’t rhyme. Or maybe it did,

If it were in another language. Except I don’t speak any other languages.²

Why did my grant crash and burn?³ How did I fail so badly?

Okay, it wasn’t an awful score, but miles away from being funded.

It’s just so unfair, this whole rejection thing. I mean, why can’t I get agood review?

Oh, but drat⁴, I hate revising grants. They eat my time like lions, or maybe

Gerbils. Or knockout mouse lines I have to list. Make a note, I have to remember

To list all the bloody mouse lines. I got dinged⁵ on that. I shouldn’t have to do this.

Yeh, lions and gerbils and knockout mice, oh my. And maybe bears.

But then there was a figure before me, to lead me on a tour, he said, of Grant Hell.

“It’s going to be a long night,” he told me, “maybe have a snack first.⁶”

“Who are you?” I asked, because he looked like Dumbledore.

And I didn’t want to go all Harry Potter on him. “I was Francis Bacon in life⁷”, said he.

Great. But hey, it could have been worse, I guess. “Can I call you Frank?”

“This is all my fault, you know,” he said, “I devised the scientific method,

which I still think is a good thing. I didn’t even think about grants.

My bad. Do you still say ‘My bad?’ Anyway, my bad.”

“But grants are a reality of how we do science. Okay, in my day we didn’t have

Grants; we had to find a benefactor.⁸ But you, at least, have a better chance.

And still, many of you would rather complain than revise your approach.”

And into Grant Hell⁹ we went. “We’re going to start at the bottom,”

He whispered, “because that’s really how it goes. Are you up for this,

or are you going to punk out?” I really wanted a drink. Not tea.

We were bathed in flames, but as shades we didn’t burn. Good, that.

“This is the Ninth Circle,” Frank said. “We won’t stay here because it’s really bad,

This is Treachery, saved for those who steal other’s projects.¹⁰” It really stank.

We passed by this one, and up to the Eighth, the circle of Fraud. It also stank.

“Some desperate sociopaths¹¹ who fake results to write a grant,” he whispered.

I was glad they had their heads firmly inserted where heads don’t properly go.

We stopped to watch Violence in the Seventh Circle. Saved for those who

Blamed their trainees for their failures. Here, the wretched watched

As their papers were shredded and labs were trashed. Serves them right.

Here in the Seventh, the trainees taunted the damned mentors.

“You could have paid attention to what I was doing!” “You jerk,

You could have mentored me, instead of dissing everything.”¹²

We climbed our way to the Sixth Circle, and here we found Heresy.

“WTF?” I asked Frank. He shrugged. “Hey, I didn’t make this place.

Here is where we go when our ideas are completely out of line with the field.”

“But that’s just good science,” I argued. “Are you telling me that these

Poor souls are here because they disagree with the common wisdom?

We have to just toe the party line and not have our own ideas?”

“No,” said Frank. “You’re missing the point. They are here because

They can’t convince anyone that their alternative viewpoint has any value.

So they huddle together and agree with themselves, and no one else.”

I approached one of the denizens. He looked at me with red-rimmed eyes.

“‘AIDS isn’t a virus’,” he quoted, “‘it is a condition of immoral lifestyle. So is cancer.’”

“I get it,” I told Frank, “alternative is fine, but stupid is stupid.”

Up to the Fifth Circle, we roamed. Here was Anger. The wretched moaned,

And complained about their fate to each other, which was pretty awful.

Since they were all working on pretty much the same things.

“It’s all your fault!” said one to the group. “If I were the only one in this field,

I’d have my grants funded. Everything I proposed was perfectly fine.”

“That’s what I said,” complained another. “Except it’s your fault.”¹³

to be continued…

↵1- Mole is a huge baby about writing grants. He calls it ‘bleeding on paper.’ The process involves putting together a plan, with supporting data, for work that is meant to have not yet been attempted, but generally must be well underway, without supporting funds. These are reviewed by ‘peers,’ other scientists who may have a passing familiarity with the research area, who meet to figure out how not to support the application. Following the almost inevitable rejection, the application is revised with more supporting data (generated without support, somehow), which results in a ‘revision,’ submitted for evisceration by another set of peers. As a consequence, a great deal of work is done without support. Once approved and funded (if that happens), the now funded scientist forgets all of the bloodshed, and generally works on something else altogether.

↵2- This is true. Including English.

↵3- (cadere a pezzi). Grants are either funded or not. If they are not funded (the most common scenario), we tend to feel that they utterly failed, and in a sense this is true. Unlike hand grenades, horseshoes and the predictions of pundits, ‘close’ does not matter with grants. This is not conventional wisdom: most will assume that ‘nearly funded’ means ‘it will get funded next time.’ Unfortunately, this does not follow. Each grant or its revision should be written as though it is the first time anyone will see it. Most often, that is the case (see Footnote 1).

↵4- merde

↵5- Ho un ragno nei pantaloni (lit. I have a spider in my pants). One never knows what a reviewer will home in on, and when we are working on a revision, we often refocus on such criticisms. However, do not be deceived: while it is important to pay attention to all criticisms, these are not ‘why’ a grant does not get funded. Do not fall into the trap of thinking that by ‘fixing’ such problems, the grant will not pay off.

↵6- Mangia un panino al burro di arachidi.

↵7- 22 January 1561–9 April 1626. English. ‘Frank’ was not only a scientist but also a philosopher, statesman, jurist, orator, and author. In addition, he worked as Attorney General and Lord Chancellor of England. He makes Mole feel exceptionally lazy.

↵8- The benefactor system was one way science was done until well into the 19th century. The only alternative was to finance the work oneself (a ghastly thought). Unlike the grant system, which relies on peer review (at least on the surface), one needed to convince a benefactor to provide the funding, and often the place in which the work was done, and funds could be withdrawn at any time. While it may seem outdated, a form of this is practiced today in the form of contracts from industry, who often seem even more pernicious than the benefactors were.

↵9- Every applicant for a research grant has experienced Grant Hell in some form. Here, though, Mole is being escorted through the nine circles of Grant Hell to learn why grants are not successful. He begins at the lowest level (unlike D.A.’s tour of real Hell) because it gets better as he goes up. But it’s all pretty awful.

↵10- It is not the case that grants fail to be funded because they were stolen from someone else. Mole simply hopes this is the case. Actually, he hopes that these people will be bitten by monkeys.

↵11- Colui che morde la scimmia (lit. he who bites the monkey)

↵12- Some investigators fail to realize that the critical interactions among laboratory workers and themselves goes two ways. Criticism is an important part of the enterprise, but if this is only negative, the morale of the laboratory declines. We are not only our laboratories’ critique, but also the cheerleaders. When we support the work of our trainees, and devote ourselves to their development, they can help us find our way out of this circle of Grant Hell. Mole is forever grateful to his own trainees, who pull together to generate supporting data for ideas he writes in his grants, and once the grant is submitted (whether or not it is supported) he works with them to develop the findings further.

↵13- It is often the case that grants are not supported because too many other people are working on the same questions, which raises issues as tohow important the questions are. This is something many applicants never take into consideration, making the error of believing that given that so many others are working on something, it must be important. But those who evaluate the proposal vary in their interests, and are often unconvinced by this argument. The answer is not to try to force others out of our area of interest, but instead, to frame our research plans in such a way that we highlight how our own work is unique. Where would the field be if you had never worked in it? Where would it be if you were not supported in what you plan to do? Channel that anger into something creative, or indeed, this circle of Grant Hell will be home.

(3 votes)

Loading...

The Regeneration Next Discovery Initiative (RNDI) is a new venture with the goals of advancing discovery research and education in the broad field of tissue regeneration, and enabling translational applications for regenerative medicine.

The Executive Director will work closely with the RNDI Director, Co-Directors, and faculty members to promote and integrate discovery research, training, and applications in the broad field of tissue regeneration.  The successful candidate will oversee the coordination of the research, recruiting, teaching, and funding missions – a critical role to help shape RNDI at Duke University.  Candidates who have a Ph.D. and postdoctoral research experience in the relevant areas of developmental biology, stem cell biology, or tissue regeneration are of particular interest.  The ideal candidate will have outstanding organizational and communication skills.  The title of this position will be based on the qualifications of the applicant, which may include appointment at the academic title of Assistant Professor. The successful applicant is not expected to develop an independent research program, but there may be opportunities for teaching.

We will review applications from now until the position is filled.  Interested applicants should submit a cover letter, curriculum vitae, summary of past research accomplishments and any administrative leadership experience, and list of at least three references to AcademicJobsOnline.org

(https://academicjobsonline.org/ajo/jobs/6097).

Questions may be directed to:

Ken Poss, Department of Cell Biology, Director, RNDI (regeneration@duke.edu)

Duke University is an Affirmative Action/Equal Opportunity Employer committed to providing employment opportunity without regard to an individual’s age, color, disability, genetic information, gender, gender identity, national origin, race, religion, sexual orientation, or veteran status.

(No Ratings Yet)

Loading...

The Regeneration Next Discovery Initiative (RNDI) is a new venture with the goals of advancing discovery research and education in the broad field of tissue regeneration, and enabling translational applications for regenerative medicine.

The Executive Director will work closely with the RNDI Director, Co-Directors, and faculty members to promote and integrate discovery research, training, and applications in the broad field of tissue regeneration.  The successful candidate will oversee the coordination of the research, recruiting, teaching, and funding missions – a critical role to help shape RNDI at Duke University.  Candidates who have a Ph.D. and postdoctoral research experience in the relevant areas of developmental biology, stem cell biology, or tissue regeneration are of particular interest.  The ideal candidate will have outstanding organizational and communication skills.  The title of this position will be based on the qualifications of the applicant, which may include appointment at the academic title of Assistant Professor. The successful applicant is not expected to develop an independent research program, but there may be opportunities for teaching.

We will review applications from now until the position is filled.  Interested applicants should submit a cover letter, curriculum vitae, summary of past research accomplishments and any administrative leadership experience, and list of at least three references to AcademicJobsOnline.org

(https://academicjobsonline.org/ajo/jobs/6097).

Questions may be directed to:

Ken Poss, Department of Cell Biology, Director, RNDI (regeneration@duke.edu)

Duke University is an Affirmative Action/Equal Opportunity Employer committed to providing employment opportunity without regard to an individual’s age, color, disability, genetic information, gender, gender identity, national origin, race, religion, sexual orientation, or veteran status.

(No Ratings Yet)

Loading...

This interview first featured in the Journal of Cell Science and is part of their interview series Cell Scientists to Watch

Philip Zegerman earned his undergraduate degree from the University of Cambridge, where he later also pursued a PhD in the lab of Tony Kouzarides at the Gurdon Institute. For his postdoctoral work, he switched fields from chromatin modifications to DNA replication, and joined John Diffley’s lab at Cancer Research UK, at Clare Hall in London. In 2009, Philip moved back to Cambridge to start his own group at the Gurdon Institute. He is an EMBO Young Investigator, and his lab is interested in how the initiation of DNA replication is regulated.

What first motivated you to become a scientist?

I have strong memories of doing experiments at school, which I think greatly influenced me. I remember one experiment very clearly, when I was probably about nine or ten. We went on a field trip and trapped and counted mice in a field. Doing experiments was really lots of fun and I was good at it. I originally thought I was going be an entomologist studying beetles because I loved insects, but when I went to university I started studying medical sciences and ended up in biochemistry and molecular biology.

Your current research focuses on the regulation of DNA replication initiation. What are the particular questions your group is pursuing at the moment?

It’s vitally important for all cells to make a perfect copy of the genome once and only once in every cell cycle. We’re studying initiation as the key regulatory event that must be strictly controlled within the cell cycle. This process has really interesting implications for how proliferation and differentiation are coordinated in large organisms. It also has important implications for diseases like cancer, where failures in replication lead to genome instability, which causes cancers, but also where DNA replication is targeted by most chemotherapies.

Which organisms do you use to answer these questions?

We use a wide range of organisms. That’s the beauty of working with replication; it’s essential for all organisms. We work on eukaryotic replication, and we study mostly budding yeast, but given our interest in whole organisms, and being at the Gurdon Institute as well, we have started to diversify and look into metazoans. We have somebody working in worms, and I’ve been doing some studies in frogs. It’s very exciting.

Is it hard to work with different model systems?

It is challenging; there are some major advantages, for example funding. If you can show funders that your process is important across organisms, that’s very good; it particularly helps if it’s in an organism that they like to fund, with relevance to disease models. There are, of course, challenges. There’s always a certain amount of time it takes to get up and running in a different system. But we’re very lucky here at the Gurdon Institute – we have lots of organisms within the building, and we have a very collaborative environment, so there’s never been any ‘activation energy’ required for moving systems and everyone’s been very helpful. It’s really the perfect environment for us to test different ideas in different systems.

Are there any new techniques that you’re adapting for your research right now?

We’re setting up a live-cell assay for replication initiation in worms. By taking advantage of the beautiful microscopy you can do in worms, with our knowledge of the molecular biology of replication initiation that we take mostly from yeast, we’re trying to set up a system in which we can visualise individual initiation events on DNA, in individual cells. If it works, then we could have the first system in which we can really study replication initiation live in a whole organism.

How have your collaborations influenced your research and do you have any advice on collaborating?

I continue to see collaborators to be really important in our work, particularly as we’re moving into different systems. Of course, collaborations aren’t without difficulties, like any project. I think the important advice that I would give to somebody is: don’t collaborate for the sake of it. You have to have a goal at the end of it and that goal has to be mutually agreed. Collaborations often lead you in exciting new ways and it’s a great way of meeting new people and networking and learning new areas of science, which I think is very important.

Many early career scientists often find that the advice given by senior scientists on how to establish a successful academic career can be outdated in the current funding climate. As someone who has established their lab relatively recently, what advice would you give?

Funding has changed so significantly since the banking crash, so young scientists have to be really aware of the pitfalls. I think my major piece of advice is that most grants are time limited. You can apply for most starting grants only in a certain window after your PhD, and if that window is approaching and you’re still a postdoc, then apply for it anyway, because that window is going to close. Even if you don’t have a firm job in place, apply for the money first and then get the job. The funders are really clear that they want people who are fresh out of their PhDs. That creates extra pressure, of course. I think when I was a postdoc I was sufficiently naïve and if I were to do it again, I would probably have applied a year earlier for most funding.

You changed fields from DNA and chromatin modification after your PhD. How did that influence your career later on?

I think that changing fields is a really undervalued part of a person’s career trajectory. When I finished my PhD, I was certain that I wanted to discover a new field. That’s why I moved into DNA replication. I really think that was an important part of my career. I enjoy the field and the questions that we have, and, of course, because I have an understanding of the chromatin field, if our questions do (and they frequently do) have implications for nucleosome remodelling or chromatin assembly, I have that experience of my PhD that allows me to understand different fields, and gives me a new angle. But having said that, because most grants are time limited, there is an expectation that postdocs will get papers out really quickly and if you change fields you can be at a disadvantage. I think that’s a real shame, because it’s important for people to cross fields and even do more than one postdoc. I think that’s a positive thing.

What are your views on the feasibility of being both a good parent and a good scientist?

Being a parent is difficult full stop! I don’t think the challenges that scientists face are different to any professional or that there’s anything particularly special about science that makes it difficult to be a parent. That’s not to say that it isn’t challenging. Of course, it’s much harder for women, because being away from the bench or the lab can be difficult if it’s for long periods of time. But that shouldn’t discourage people from having children; I had both my kids when I was a postdoc, so if you’re organised then it shouldn’t be a problem – it can be managed. Lots of people do it, so it’s obviously possible.

Are universities providing enough support for scientists with families?

There’s always more support that could be given. I think that one of the major challenges in the southeast of England is the extreme pressure on nursery places that are very expensive. Universities are increasingly encouraged to provide their own childcare – the University of Cambridge has a very small number of such places. Childcare is very expensive, so any kind of incentive that can allow parents to pay their nursery fees out of their gross salary should be encouraged.

I asked you before why you became a scientist. What motivates you now?

Science is inspiring. I think that’s what motivates me – doing science that can transcend the normal desk jobs of this world, to really be inspired by something new, interesting and exciting. I also often get very inspired by going and hearing other people talk. Last week Professor Johannes Walter came from Harvard, and he gave such an excellent, exciting and interesting talk about replication regulation that not only was I impressed by his work, but it encouraged me to go back to the lab and find out something that could be as good as that, and I find that very motivating.

Could you share with us an interesting fact about yourself that people wouldn’t know just by looking at your CV?

I do a lot of gardening, and I grow lots of fruit and vegetables. We had a very big garden when I was a child, and I was in charge of mowing the lawn and doing the gardening, and it stayed with me. I had an allotment even when I lived in London. I’m still fascinated by plants and insects, and growing things, and my kids are quite into it now. It’s becoming a family hobby. They like eating the food more than growing it though.

Also watch this additional clip:

(2 votes)

Loading...