Posts Tagged ‘mouse ES cells’

New ways of inducing pluripotency and additional applications for the CRISPR-Cas system

Saturday, February 1st, 2014
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The CRISPR-Cas system for genome editing was launched in 2013 for applications in animal transgenesis and continues advancing in 2014

The CRISPR-Cas system for genome editing was launched in 2013 for applications in animal transgenesis and continues advancing in 2014

The first weeks of 2014 have generated interesting technical advances in animal transgenesis, and prestigious ISTT members have been involved in them. If this is just a sample of what will come next it would seem appropriate to call this starting 2014 year the wonder year. This past week we knew about a new manner for inducing pluripotency, simply exposing somatic cells to a low pH, using a physical stimulus, transiently applied during a short period of time. This acidic exposure appears to trigger the reprogramming steps required to convert somatic into fully capable pluripotent cells, sustenting the generation of germ-line transmitting chimeras. Furthermore, these STAP (Stimulus-Triggered Acquisition of Pluripotency) cells appear to be able to contribute to both the embryonic and extra-embryonic lineages, thus constituting a unique status of pluripotency.  These awesome two papers, by Haruko Obokata and collaborators, have been published in Nature, and include as co-author in one and senior corresponding author in the other, ISTT member Teruhiko Wakayama, the first scientist awarded the ISTT Prize.

Also last week we learnt about the first non-human knockout primates. A group of Chinese scientists (Yuyu Niu and collaborators), including the most prestigious centres involved in the generation of animal models in China, published a paper in Cell where they reported a new application for the powerful and novel CRISPR-Cas technology to produce mutant monkeys. They generated, for the first time, twin cynomolgus monkeys (Macaca fascicularis) with two targeted loci, Ppar-g and Rag1, in one single step. This collaborative work included as co-authors ISTT member and ISTT Prize awarded scientist Qi Zhou, as well as Xiaoyang Zhao, who received the first ISTT Young Investigator Award.  This achievement, which was not possible to date with standard technologies, illustrates the unlimited power of the CRISPR-Cas system.

We first learnt about the CRISPR-Cas system, as the responsible for adaptative bacterial immunity,  in mid 2012. But it was not until last year, 2013, when the molecular reagents become amenable and applicable for genome editing in animal cells and embryos, for the generation of a variety of genetically-modified animals, including all sorts of transgenic and mutant types, with an explosion of papers and applications. Today, 1st February 2014, as many as 88 papers appear listed in PubMed combining “CRISPR genome editing”. The amazing simplicity of this sytem, and the ease by which anyone can start using this technology in the lab, simply obtaining the two required plasmids (carrying the RNA guide, where the target homologous sequence must be engineered, and the Cas9 nuclease) from diverse providers, including Addgene, explains why the CRIRPR-Cas technology is now being considered a true revolution in our field, in animal transgenesis.

Transgenic mice obtained from androgenetic haploid embryonic stem cells

Wednesday, October 3rd, 2012
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Transgenic mice obtained from androgenetic haploid embryonic stem cells (W Li et al. Nature 000, 1-5 (2012) doi:10.1038/nature11435)

Transgenic mice obtained from androgenetic haploid embryonic stem cells. Image adapted by permission from Macmillan Publishers Ltd:NATURE (W Li et al. Nature 000, 1-5 (2012) doi:10.1038/nature11435), copyright (2012).

Prof. Xiao-Yang Zhao (ISTT Ordinary member and awarded the first ISTT Young Investigator Award in Florida (USA), at the TT2011 meeting) and Prof. Qi Zhou (ISTT Honorary member and awarded the third ISTT Prize in Uppsala (Sweden), at the TT2004 meeting), and his colleagues from the State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, in Beijing (China), as well as other collaborating institutions, just published a letter in Nature describing how live transgenic mice can be obtained from androgenetic haploid embryonic stem cells (Li et al. Nature 2012).

Androgenetic haploid embryonic stem cells produce live transgenic mice
Wei Li, Ling Shuai, Haifeng Wan, Mingzhu Dong, Meng Wang, Lisi Sang, Chunjing Feng, Guan-Zheng Luo, Tianda Li, Xin Li, Libin Wang, Qin-Yuan Zheng, Chao Sheng, Hua-Jun Wu, Zhonghua Liu, Lei Liu, Liu Wang, Xiu-Jie Wang, Xiao-Yang Zhao & Qi Zhou
Nature (2012) doi:10.1038/nature11435.

Mouse androgenetic haploid embryonic stem (ahES) cells can be established by transferring sperm into an enucleated oocyte. These ahES cells maintain haploidy and are stable in culture. In addition, these ahES cells can contribute to the germline of chimeric mice when microinjected into blastocysts. Furthermore, these ahES cells can be delivered by intracytoplasmic injection into mature oocytes, eventually resulting into viable fertile mice that will inherit any genetic modification previously transferred to ahES cells while in culture. As stated by the authors, this work “provides a new approach for genetic manipulation in animal models without available germline-competent ES cells, including non-human primates, as modifications in such haploid stem cells could be transmitted to offspring through intracytoplasmic injection into mature oocytes, which may serve as a more efficient and simple strategy for gene-targeting studies“.

Earlier this year, in the April 27 issue of Cell, an independent study, developed in parallel, from Guo-Liang Xu’s (State Key Laboratory of Molecular Biology) and Jinsong Li‘s (State Key Laboratory of Cell Biology) laboratories from the Institute of Biochemistry and Cell Biology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences in Shanghai (China) and their collaborators, including Xiang Gao (Model Animal Research Center, Nanjing University, Nanjing, China), reported similar extraordinary findings.

Generation of genetically modified mice by oocyte injection of androgenetic haploid embryonic stem cells.
Yang H, Shi L, Wang BA, Liang D, Zhong C, Liu W, Nie Y, Liu J, Zhao J, Gao X, Li D, Xu GL, Li J.
Cell. 2012 Apr 27;149(3):605-17.

These three scientists, Qi Zhou (Beijing, China), Jinsong Li (Shanghai, China) and Xiang Gao (Nanjing, China) will participate as invited speakers in the next 11th Transgenic Technology meeting (TT2013) that will be held in Guangzhou (China), on 25-27 February 2013. In particular, both Qi Zhou and Jinsong Li will be presenting their most recent and excellent works on the isolation of androgenetic haploid embryonic stem cells and their use to produce genetically-modified mice.

Once again, the International Society for Transgenic Technologies (ISTT) is pleased to present the latest advances in animal transgenic technology at the TT meetings, next one (TT2013) to be held in Guangzhou (China), in February 2013. Anyone interested in these transgenic techniques and their applications should not miss this great opportunity to learn all these new developments, directly presented by their inventors.

Course on Genetic Manipulation of ES Cells, Hinxton, Cambridge, UK, 5-18 November 2012

Monday, March 19th, 2012
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Course on Genetic Manipulation of ES Cells

Course on Genetic Manipulation of ES Cells

Course on Genetic Manipulation of ES Cells, 5-18 November 2012
Wellcome Trust Genome Campus, Hinxton, Cambridge, UK
Deadline for applications: 6 July 2012

Course summary
This laboratory-based training course will provide a comprehensive overview and practical laboratory experience of the genetic manipulation of mouse ES cells for a broad range of applications. Recent advances in genome informatics, recombineering, transposon technology and uses of conditional gene targeting will be covered in lectures by both instructors and invited expert speakers and through interactive demonstrations.Laboratory work will focus on the culture and transfection of ES cells, design and construction of gene targeting vectors from BACs by recombineering, genotyping of gene targeting events and the practical use of transposon technology. Participants will also be trained in the informatics and practical use of public gene targeting resources being produced by the IKMC (International Knockout Mouse Consortium).

Course instructors
Professor Francis Stewart (Dresden University of Technology, Dresden, Germany)
Dr William Skarnes (Wellcome Trust Sanger Institute, Cambridge, UK)
Dr Pentao Liu (Wellcome Trust Sanger Institute, Cambridge, UK)
Dr Barry Rosen (Wellcome Trust Sanger Institute, Cambridge, UK)

The Course programme includes the following topics:

1. Informatics
The informatics underlying the visualization of gene structures and the design of gene targeting vectors, recombineering oligos and genotyping primers will be demonstrated. Students will also run their own gene targeting designs using web-based tools.The informatics of locating existing public IKMC gene targeting resources will also be covered.

2. Recombineering of Gene Targeting Vectors
Students will build a variety of targeting constructs from BACs using recombineering technology. The theoretical principles underlying both recombineering and rational targeting vector design will be emphasized by both lectures and practical exercises.

3. ES Cell Culture
Students will learn feeder-dependent and feeder-free culture of ES cells derived from 129 and C57BL/6 mouse strains.ES cell colonies will be picked, expanded and frozen and subsequently thawed to test their integrity.

4. Gene Targeting in ES Cells
Students will electroporate conditional gene targeting constructs into ES cells and learn to genotype cells by LR-PCR and qPCR-based methods.

5. Transposon Technology
Students will be introduced to the uses of the highly efficient piggyBac transposon system for expression, mutagenesis and mouse induced pluripotent stem cell (iPS) cell generation.

6. Modular Targeting Resources
Students will assemble a variety of modular knock-in targeting vectors from IKMC resources and analyse their integrity.Recombination Mediated Cassette Exchange (RMCE) using Flp and Cre recombinases will be used to modify IKMC conditional alleles directly in ES cells.

Additional information and instructions to apply are available from the web site of this course

Allan Bradley will be awarded the 9th ISTT Prize at the TT2013 meeting

Tuesday, January 31st, 2012
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Allan Bradley will be awarded the 9th ISTT Prize at the TT2013 meeting (Picture kindly provided by WTSI)

Allan Bradley will be awarded the 9th ISTT Prize at the TT2013 meeting (Picture kindly provided by WTSI)

The International Society for Transgenic Technologies (ISTT) is pleased to announce that Professor Allan Bradley, Director Emeritus of the Wellcome Trust Sanger Institute (WTSI), and leader of the Mouse Genomics Team at WTSI, will be awarded the next (9th) ISTT Prize for his outstanding contributions to the field of transgenic technologies. Professor Allan Bradley will receive the award at the next Transgenic Technology meeting (TT2013), which will be held in Guangzhou (PR China) on February 25-27, 2013. This award has been agreed upon by the ISTT Prize Committee, consisting of the ISTT President and Vice-President, the CEO of genOway, as the company generously sponsoring the award, and the previous ISTT Prize awardees.

In awarding this prize to Dr. Bradley, the ISTT Prize committee acknowledges his many fundamental contributions to the science and technology of manipulating the mouse genome. His pioneering mouse embryonic stem (ES) cell work in the 1980s, demonstrating germ-line transmission and the great potential of ES cells to generate mice carrying mutations in endogenous genes, established milestones in a field that saw the award of the 2007 Nobel Prize in Physiology or Medicine to Mario Capecchi, Martin Evans, and Oliver Smithies. Later, Dr. Bradley generated a number of broadly relevant knockout mouse models (for example p53, Rb, Wnt-1) that are still used regularly today. His subsequent research has developed new methods for the genetic analysis and genetic modification of mice (such as chromosome engineering and the functional genetic analysis of mouse chromosome 11). These developments have been instrumental for advancing mouse genetics studies and the use of mice to understand the human genome. Furthermore, his strong vision and leadership at the Wellcome Trust Sanger Institute, which he directed from 2000-2010, was key to creating the EUCOMM/KOMP (IKMC) initiative to systematically disrupt every gene in the mouse genome, resulting in a massive impact on the field of transgenic technologies.

Dr. Bradley received his BA, MA and PhD in Genetics from the University of Cambridge. His PhD studies in Martin Evans’ laboratory, completed in 1984, laid the foundation for making knockout mice.

In 1984 Allan Bradley and Liz Robertson demonstrated that ES cells could be transmitted through the germ-line of mice (Bradley et al. 1984, Nature) and two years later reported that ES cells could be used to generate mice with mutations in endogenous genes (Kuehn, Bradley et al. 1987, Nature).

In 1987, he took an appointment as Assistant Professor at Baylor College of Medicine, Houston, Texas. He was named as a Howard Hughes Medical Institute Investigator in 1993 and was promoted to full Professor in 1994. At Baylor, his laboratory played a seminal role in developing the methods, technology and tools for genetic manipulation in the mouse. As a result, mice can now be generated with changes as subtle as an alteration in a single nucleotide or as massive as the deletion, duplication or inversion of millions of base pairs with a technology now known as chromosome engineering (Ramirez-Solis et al. 1995, Nature). The Bradley laboratory used ES cell technology extensively, generating and analysing many of the first generation of mouse knockouts (McMahon & Bradley, 1990, Cell; Donehower et al. 1992, Nature; Lee et al. 1992, Nature; Jones et al. 1995, Nature; Sharan et al. 1997, Nature) while helping numerous other laboratories to utilize this technology. This work has provided key functional information for many genes with an emphasis on cancer, DNA repair and embryonic development. While at Baylor, Bradley created the Mouse Club, originally consisting of meetings among his, Phil Soriano’s, and Gerard Karsenty’s laboratories, with Richard Behringer’s lab joining later. The Mouse Club has since expanded and has been meeting for more than two decades, every Tuesday afternoon. Richard Behringer remembers: “I was always impressed that Allan would always attend the Mouse Club if he was in town. He put training of students and postdocs as a high priority.”  Many former colleagues and trainees from Allan Bradley’s lab have fond memories of his mentoring and support. Y. Eugene Yu says: “When tropical storm Allison flooded Houston and the Texas Medical Center in 2001, which killed 90,000 research animals, Allan flew in from UK immediately after the news broke. He personally led the effort in the devastated animal facility to rescue the remaining precious mouse strains, many of which were unique.”

In November 2000, Allan Bradley returned to the United Kingdom as Director of the Sanger Centre, now called the Wellcome Trust Sanger Institute (WTSI), shortly after the first draft of the human genome sequence was released. In 2002, he oversaw the publication of the mouse genome sequence.

His 10-year plan for the WTSI aimed to transform it from a centre that just sequences DNA to one that studies the biology of sequences. First focussing on creating the required physical infrastructure, then diversifying the institute’s research, his efforts have helped turn the WTSI into an international hub of science, with strong programs in human genetics, informatics, pathogen genetics, and model organisms. Among other projects in which the WTSI plays a leading role is the largest systematic gene knockout project ever attempted in ES cells, funded by the European Union (EUCOMM) and National Institutes of Health (KOMP), and now coordinated by the International Knockout Mouse Consortium (IKMC) (Skarnes et al., 2011 Nature; Prosser et al., 2011 Nature Biotechnology).

Under Allan Bradley’s direction, the WTSI has become a reference centre for outreach activities, offering teaching resources, welcoming students for work placements and promoting school visits that broadcast scientific achievements to the general society; in his words “inspiring the next generation of scientists.”

Allan Bradley has also been very active in commercializing the numerous novel methods his laboratory has developed over the years to engineer the genomes of mice. He is registered as a co-inventor on more than 25 patents and is a founder and Chief Scientific Officer of Kymab Limited. He also co-founded several companies, including GenPharm International and Lexicon Genetics in 1995. Using proprietary gene trapping and gene targeting technologies, Lexicon Genetics created the world’s first large repository of genetically modified mouse embryonic stem cells, known as OmniBank, and established a large-scale program to discover the physiological and behavioural functions of mammalian genes, with almost 5,000 gene knockouts studied in mice. This effort is now being extended to the entire mouse genome in a public domain project coordinated by the International Mouse Phenotyping Consortium (IMPC) and funded by NIH in the USA and other funding bodies internatioanlly.

In July 2002 Allan Bradley was elected a Fellow of the Royal Society. He has authored over 260 scientific articles and book chapters and currently runs an active research group that is exploring gene function on a large scale and continues to develop new tools and technologies for mouse genetics research.

The ISTT Prize: a piece of art created by Bela Rozsnyay

The ISTT Prize: a piece of art created by Bela Rozsnyay

Allan Bradley will join in 2013, at the TT2013 meeting, the following group of outstanding scientists previously awarded the ISTT Prize:

Ralph L. Brinster, ISTT Prize, TT2011, Florida, USA, Prize Announcement, Award ceremony
A. Francis Stewart, ISTT Prize, TT2010, Berlin, Germany, Prize Announcement, Award ceremony
Brigid Hogan, ISTT Prize, TT2008, Toronto, Canada, Prize Announcement, Award ceremony
Charles Babinet, TT2007, Brisbane, Australia, Obituary (1939-2008)
Andras Nagy, TT2005, Barcelona, Spain
Qi Zhou, TT2004, Uppsala, Sweden
Kenneth C. McCreath, TT2002, Munich, Germany
Teruhiko Wakayama, TT2001, Stockholm, Sweden

Download this Award Announcement as a ISTT Press Release Document

NIH Course: TRAC 12:Transgenic Technology: Methods and Application, Jan 9-13, 2012

Saturday, December 3rd, 2011
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TRAC 12:Transgenic Technology: Methods and Application
TRAC 12:Transgenic Technology: Methods and Application

The TRAC 12:Transgenic Technology: Methods and Application Course is a four-day Biotechnology Training Courses at the National Institutes of Health that will be held on January 9-12, 2012. According to its web site: “This lecture/laboratory demonstration course is intended for those who wish to develop a strong background in transgenic techniques and applications. The two dominant approaches to transgenic technology will be emphasized, namely classic transgenic production by pronuclear microinjection and knockout transgenic production(gene targeting) by blastocyst injection with embryonic stem (ES) cells. The comprehensive coverage will address supporting technology (e.g. cryopreservation, rederivation, Intracytoplasmic Sperm Injection(ICSI), In Vitro Fertilization (IVF) , animal husbandry and surgery) as well as the molecular biology/recombinant DNA aspects of transgenesis.”

The TRAC12 Course will cover the following topics: Transgenic Technology overview  (Embryonic Development,  Optimizing Constructs for in vivo Expression,  Preparation of Females for Embryo Collection, Pronuclear Microinjection,  Generation of Pseudopregnant Females,  Implantantation in Foster Mothers,  Identification of Transgenic Progeny,  Optimization of Breeding Population from Founders);  Lentiviral and BAC transgenisis;  Gene Targeted Transgenisis (Isolation and Maintenance of Totipotent embryonic Stem Cells for the Generation of Chimeric Mice; Homologous Recombination in ES Cells; Construct Design, Transfection, Selection; Generation of Gene Knockout Mice using Targeted ES Cells; Blastocyst Injection; Optimizing Germ-line Chimeras);  Alternative Methods for Generating Targeted Mice: 8_cell Embryo Injection and ES Cell Aggregation;  transgenic phenotype analysis;  Transgenic Animal Model Supporting Techniques (Cryopreservation and rederivation;  ICSI and IVF; Transgenic Model Future Prospects); Transgenic Model FuFuture Prospects; Sampling Project Question and Answer; Live and Video Demonstrations and hands on work (e.g. microinjection; Superovulation; Embryo Harvest and Transfer).

Applications to attend this TRAC 12 course can be submitted through this web site.

Published in Nature: A conditional knockout resource for the genome-wide study of mouse gene function

Thursday, June 16th, 2011
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Targeting strategies and constructs used by KOMP-CSD and EUCOMM

Targeting strategies and constructs used by KOMP-CSD and EUCOMM

Today’s issue of Nature includes the article reporting the great initiative, efforts and results achieved so far by the International KnockOut Mouse Consortium (IKMC) towards the systematic gene-targeting of the mouse genome, aiming to functionally annotate and thus deciphering the precise role of all genes encoded by a mammalian genome (mouse), most similar to the human genome. The authors are members of the KOMP (the National Institutes of Health Knockout Mouse Program) and EUCOMM (the European Conditional Mouse Mutagenesis) international projects.

In this work, the authors report the establishment of a high-throughput gene-targeting efficient strategies and a successful pipeline to produce reporter-tagged, conditional alleles on an unprecedent scale. As they report, “more than 12,000 vectors and 9,000 conditional targeted alleles“ have been produced so far. Targeted ES cells and targeting vectors are available from KOMP and EUMMCR. Mice derived from EUCOMM ES cells are available as live animals or cryopreserved embryos from EMMA.

A conditional knockout resource for the genome-wide study of mouse gene function
William C. Skarnes, Barry Rosen, Anthony P. West, Manousos Koutsourakis, Wendy Bushell, Vivek Iyer, Alejandro O. Mujica, Mark Thomas, Jennifer Harrow, Tony Cox, David Jackson, Jessica Severin, Patrick Biggs, Jun Fu, Michael Nefedov, Pieter J. de Jong, A. Francis Stewart & Allan Bradley.  Nature 474 (337-342) Date published: 16 June 2011 DOI: doi:10.1038/nature10163

Additional comments available here.

WTSI Course on Genetic Manipulation of ES cells

Tuesday, June 14th, 2011
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WTSI Course on Genetic Manipulation of ES cells, 31 October-13 November 2011

WTSI Course on Genetic Manipulation of ES cells, 31 October-13 November 2011

The Wellcome Trust Sanger Institute (WTSI), at Hinxton (UK), organizes a course on “Genetic Manipulation of ES cells that will be held on 31 October-13 November 2011. This Wellcome Trust Advanced Course will provide a comprehensive overview and practical laboratory experience of the genetic manipulation of mouse ES cells for a broad range of applications. The course instructors include Course instructors Prof. Francis Stewart, ISTT Honorary Member, from Dresden University of Technology, Germany and Dr William Skarnes, Dr Pentao Liu, and Dr Barry Rosen, from WTSI, UK.

Guest speakers include Toni Cathomen (Hannover Medical School, Germany), Austin Smith (Wellcome Trust Centre for Stem Cell Research, UK), Roger Pedersen (University of Cambridge, UK), Allan Bradley (Wellcome Trust Sanger Institute, UK), Meinrad Busslinger (Research Institute of Molecular Pathology, Austria) and Timm Schroeder (Helmholtz Zentrum München, Germany). Application deadline: 8 July 2011.

A brochure for this course can be downloaded from here.

Advanced Protocols for Animal Transgenesis. An ISTT Manual (Due: August 2011)

Saturday, April 9th, 2011
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Advanced Protocols for Animal Transgenesis. An ISTT Manual. Shirley Pease & Thomas L. Saunders (eds.), Springer 2011, 1st edition (Due: August 2011)

Advanced Protocols for Animal Transgenesis. An ISTT Manual. Shirley Pease & Thomas L. Saunders (eds.), Springer 2011, 1st edition (Due: August 2011)

The International Society for Transgenic Technologies (ISTT), in collaboration with Springer, is pleased to announce the next publication of the book entitled “Advanced Protocols for Animal Transgenesis. An ISTT Manual“, edited by Shirley Pease and Thomas L. Saunders, whose 1st edition is expected to be published by August 2011.

This laboratory manual, published by Springer in cooperation with the International Society for Transgenic Technology (ISTT), provides almost all current methods that can be applied to the creation and analysis of genetically modified animals. The chapters have been contributed by leading scientists who are actively using the technology in their laboratories, most of them members of the ISTT. Based on their first-hand experience the authors also provide helpful notes and troubleshooting sections.

Topics range from standard techniques, such as pronuclear microinjection of DNA, to more sophisticated and modern methods, such as the derivation and establishment of embryonic stem (ES) cell lines, with defined inhibitors in cell culture medium. In addition, related topics with relevance to the field are addressed, including global web-based resources, legal issues, colony management, shipment of mice and embryos, and the three R’s: refinement, reduction and replacement.

Table of contents:

  • Karen S. Canady: Patent and licensing issues in transgenic technology.
  • Lluís Montoliu: Global Resources: Including Gene Trapped ES Cell Clones: Is Your Gene Already Knocked Out?.
  • Eduardo Moltó, Cristina Vicente-García and Lluís Montoliu: Designing Transgenes for Optimal Expression.
  • Thomas L. Saunders: Gene Targeting Vector Design for Embryonic Stem Cell Modifications.
  • Thomas J. Fielder and Lluis Montoliu: Transgenic Production Benchmarks.
  • Katja Becker and Boris Jerchow: Generation of Transgenic Mice by Pronuclear Microinjection.
  • Séverine Ménoret, Séverine Remy, Laurent Tesson, Claire Usal , Anne-Laure Iscache  and Ignacio Anegon: Generation of Transgenic Rats using Microinjection of Plasmid DNA or Lentiviral vectors.
  • Almudena Fernández, Diego Muñoz and Lluís Montoliu: Generation of Transgenic Animals by Use of YACs.
  • Michael G. Zeidler, Margaret L. Van Keuren and Thomas L. Saunders: BAC Transgenes, DNA Purification, and Transgenic Mouse Production.
  • Carlos Lois: Generation of Transgenic Animals with Lentiviral Vectors.
  • Aron Geurts, Lajos Mates and Darius Balciunas: Vertebrate Transgenesis by Transposition.
  • Karen M. Chapman, Dalia Saidley-Alsaadi, Andrew E. Syvyk, James R. Shirley, Lindsay M. Thompson and F. Kent Hamra: Rat Spermatogonial Stem Cell Mediated Gene Transfer.
  • Sayaka Wakayama, Nguyen Van Thuan and Teruhiko Wakayama: Mouse Cloning by Nuclear Transfer.
  • Elizabeth D. Hughes and Thomas L. Saunders: Gene Targeting in Embryonic Stem Cells.
  • Wojtek Auerbach and Anna B. Auerbach: The Importance of Mouse ES Cell Line Selection.
  • Marina Gertsenstein: Tetraploid Complementation Assay.
  • Elizabeth Williams, Wojtek Auerbach, Thomas M. DeChiara and Marina Gertsenstein: Combining ES cells with Embryos.
  • Kristina Nagy and Jennifer Nichols: Derivation of Murine ES Cell Lines.
  • Ping Li, Eric N Schulze, Chang Tong and Qi-Long Ying: Rat Embryonic Stem Cell Derivation and Propagation.
  • Han Li, Katerina Strati, Verónica Domínguez, Javier Martín, María Blasco, Manuel Serrano and Sagrario Ortega: Induced pluripotency: generation of iPS cells from mouse embryonic fibroblasts.
  • Anna B. Auerbach, Peter J. Romanienko and Willie H. Mark: The Preparation and Analysis of DNA for Use in Transgenic Technology.
  • Karen Brennan: Colony Management.
  • Belen Pintado and Juan Hourcade: Cryopreservation.
  • Shirley Pease: Shipment of Mice and Embryos.
  • Jorge M. Sztein, R.J. Kastenmayer and K.A. Perdue: Pathogen Free Mouse Rederivation by IVF, Natural Mating and Hysterectomy.
  • Jan Parker-Thornburg: Refinement, Reduction and Replacement

ISTT Members are entitled to a 33% discount on the book price.

B6N ES cells can be aggregated with albino outbred ICR morulas for the efficient production of chimeras

Friday, July 16th, 2010
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ES cells set for aggregation with a mouse morula

ES cells set for aggregation with a mouse morula

Marina Gertsenstein (member of ISTT) and her colleagues, from the Samuel Lunenfeld Research Institute, Mount Sinai Hospital and the Toronto Centre for Phenogenomics, in Toronto (ON, Canada), have recently published a scientific article in PLoS ONE describing the efficient generation of mouse chimeras with C57BL/6N ES cells by aggregation with standard albino outbred ICR morulas. As detailed in this work, the use of chemically-defined ES cell culture medium (2i) appears to be crucial for the success of the experiment. This is an interesting development that can help the biomedical research community for the easy and rapid generation of C57BL/6N -derived chimeras at a reduced cost. It also complements the toolbox by which chimeras can be obtained with the popular C57BL/6N ES cells, used in the large-scale international consortiums (i.e. IKMC) aiming to generate systematic knock-outs of all genes in the mouse genome.

Origin of mouse ES cell lines

Monday, December 8th, 2008
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Mouse embryonic-stem cells
Mouse embryonic-stem cells

There are many mouse embryonic stem (ES) cells commonly used and available. Most of them are derived from any of the 129 mouse sub-strains. If you would like to know the mouse strain of origin of the mouse ES cell you are using you can browse the document released by The Jackson Lab a few years ago (1999, 2001) on “New 129 nomenclature – revised” or, better, download the entire list of available mouse ES cells according to their common name and mouse strain of origin, prepared and served from the Mouse Genome Informatics ftp WEB server.


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