7th Workshop on Innovative Mouse Models (IMM2013), Leiden, The Netherlands, 13-14 June 2013

7th Workshop on Innovative Mouse Models (IMM2013), Leiden, The Netherlands, 13-14 June 2013
7th Workshop on Innovative Mouse Models (IMM2013), Leiden, The Netherlands, 13-14 June 2013

The International Society for Transgenic Technologies (ISTT) has agreed to co-sponsor the 7th Workshop on Innovative Mouse Models (IMM2013), which will be held at the Leiden University Medical Center (LUMC), in Leiden, The Netherlands, on 13-14 June 2013. This will be the third consecutive edition of this meeting series that is granted with the co-sponsorship of the ISTT, after the IMM2011 and the IMM2009 editions, due to its interest and relevance for the community of researchers using genetically-modified mice. The IMM2013 workshop is organized by: Jos Jonkers (NKI-AVL, Amsterdam, The Netherlands), Paul Krimpenfort (NKI-AVL, Amsterdam, The Netherlands), Werner Mueller (University of Manchester, Manchester, United Kingdom), Hein te Riele (NKI-AVL, Amsterdam, The Netherlands), Els Robanus-Maandag (LUMC, Leiden, The Netherlands), Marian van Roon (VU, Amsterdam, The Netherlands and ISTT Member) and Sjef Verbeek (LUMC, Leiden, The Netherlands and ISTT Member).

According to the IMM2013 workshop web page: The primary goal of this two-day workshop is to bring together a diverse group of scientists interested in advanced genome alteration approaches in the mouse, including key developers of emerging technologies as well as researchers who wish to apply and assess these new approaches. The IMM2013 workshop will encourage an in-depth and unvarnished discussion of these technologies and novel developments. This 2-day workshop will have a mixture of invited speakers and selected presentations. Keynote lectures will be given by:

  • Anton Wutz (UK) haploid ESC
  • Haoyi Wang (US) iPS / TALENs
  • Bill Skarnes (UK) high throughput TALENs
  • Ben Davis (UK) docking site/ new recombinases
  • Yann Herault (France) Cre transgenic mice
  • Kevin Brindle (UK) MRI
  • Mathijs Verhagen (NL) large scale phenotyping
  • B Kappes (US) TALENs
  • Zoltan Ivics (Germany) Transposons

ISTT survey on production of transgenic mice has been completed

ISTT survey on production of transgenic mice has been completed
ISTT survey on production of transgenic mice has been completed

On November 1st, 2008, the International Society for Transgenic Technologies (ISTT) launched the ISTT survey on production of transgenic mice. This survey was presented by Tom Fielder (Univ. California-Irvine, USA) at the TT2008 meeting (Toronto, Canada) where we shared the outcome of a pilot survey conducted with a limited number of transgenic facilities. The purpose of this survey was to establish performance standards for the production of transgenic mice by pronuclear microinjection of DNA. Our aim was to benefit all transgenic facilities by establishing officially recognized standards for such measures as embryo and pup yields, rates of transgenesis, etc., as well as the expected variability in such measures. Existing facilities would then be able to refer to independent standards in dealing with dissatisfied clients, and new facilities would have achievable goals to aim for. We hoped that it might even be possible to discover an optimum combination of factors that would allow every facility to improve. Our goal was to obtain data from at least 100 different facilities world-wide. However, in September 2009, when the survey was closed, we had received raw microinjection data from only 66 independent facilities, including more than 17,000 microinjection sessions. Preliminary global analyses from merged submitted data were presented by Tom Fielder at the TT2010 meeting (Berlin, Germany) and, some of these analyses, were reported in a book chapter on “Transgenic Production Benchmarks” from the ISTT Manual: Advanced Protocolos for Animal Transgenesis (S. Pease & T.L. Saunders, 2011; Springer). During 2011 we began to produce the announced customized statistical analysis for each participating facility. This proved to be a very demanding and time-consuming task. An example of such a customized analysis was presented by Tom Fielder at the TT2011 meeting (Florida, USA).

Eventually, in September 2012, all facility reports were sent to their original contributors. We certainly appreciate the fact that the analyses would have been more valuable had they been completed sooner. At the same time, we hope that contributing facilities will appreciate the enormity of this project, which has obviously required much more effort than we anticipated. We would like to acknowledge the help of our collaborator, Laura Barrios, professional statistician at CSIC, whose contributions were critical to achieving the aims of this survey. Without her expert guidance, provided free of charge, the survey could not have happened.

Now, that the ISTT Survey on production of transgenic mice has been completed, we will prepare summary publications discussing in greater details the main findings and conclusions. Also, we are pleased to announce the launching of a new resource, related to this transgenic survey. We are now accepting additional microinjection data from facilities that wish to compare their performance against the survey’s original data set (66 facilities/>17,000 microinjection sessions). Interested facilities are encouraged to visit this new resource page.

Tom Fielder & Lluís Montoliu

Transgenic mice obtained from androgenetic haploid embryonic stem cells

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.

What are we talking about in the transgenic-list?

What are we talking about in the transgenic-list?
What are we talking about in the transgenic-list?

After 16 years and more than 20,000 messages posted one could ask: what are we talking about in the transgenic-list? What are the topics mostly discussed in this very popular list among people interested in animal transgenesis? One way to address this question is by using Wordle, a program that analyzes a text and extract the words that are mostly used and draws a graphic where the most frequent words appear more prominently.

Download a WORDLE for the transgenic-list (tg-l): small, big, huge.

What you see above is the result of providing Wordle with the subjects from these more than 20,000 messages posted through the tg-l over the past 16 years. Watching the resulting graphic provides a rather clear answer: we appear to be mostly talking about ES cells, mice, embryos, transgenic, injection, IVF, DNA, etc… but clearly, ES cells is the most popular topic at the tg-l. Intreresting. A simple but very informative manner to present the usage of words in subjects of messages posted through the transgenic-list.

World Map of Transgenic Core Facilities

World Map of Transgenic Core Facilities
World Map of Transgenic Core Facilities

At the International Society for Transgenic Technologies (ISTT) web site, according to the aims of our Society. we care to provide the entire scientific community with as much information as possible regarding how and where to generate genetically modified animals, particularly transgenic and knockout mice, as useful animal models for research projects in biology, biotechnology and biomedicine. One of these resources of information is the World Map of Transgenic Core Facilities, currently holding more than 125 links to web sites of transgenic core facilities located in 27 countries, world wide. The transgenic core facilities can be easily found in a list, arranged per country, or using a useful Google Maps built-in feature depicting the geographic location of each transgenic facility.

Is your transgenic core facility not yet listed in the World Map of Transgenic Core Facilities offered from the ISTT web site? No problem. Whether public or private, whether based on an academic environment or associated with a company, all transgenic core facilities, all initiatives meant to produce transgenic animals (mice, rats, other mammals, other vertebrates,…) on demand, for research purposes, are welcome and we, at the ISTT, will be pleased to include all these links in our web site. Please contact us at istt@transtechsociety.org and send us your web link and contact details of your transgenic core facility and we will be more than happy to add your transgenic core facility to the list of World Map of Transgenic Core Facilities.

Thanks for submitting the web site of your Transgenic Core Facility to the ISTT.

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

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.

Publication of the Academy of Medical Sciences Report on Animals containing human material

Publication of the Academy of Medical Sciences Report on Animals containing human material
Publication of the Academy of Medical Sciences Report on Animals containing human material
The Academy of Medical Sciences has published the report of a working group study on ‘animals containing human material (ACHM)’ today. The report was prepared by a working group, chaired by Professor Martin Bobrow CBE FRS FMedSci. The working group of experts included Prof. Robin Lovell-Badge (NIMR-MRC, UK), member of the International Society for Transgenic Technologies (ISTT). The Academy of Medical Sciences promotes advances in medical science and campaigns to ensure these are translated into healthcare benefits for society. The report examines the use of ACHM from scientific, social, ethical, safety and regulatory perspectives, and highlights how ACHM are used both in investigational research, and in the development and production of therapeutics. The study was informed by open call for evidence, expert evidence, and a commissioned public dialogue.
The report concludes that the majority of ACHM research does not pose ethical or regulatory difficulties, but identifies 3 areas that will need careful oversight in future:
  • Modification of an animal’s brain which might lead to human-like cognition;
  • Changing an animal so it has human appearance or characteristics (e.g. skin, facial or limb features, speech); and the
  • Development of human-derived sperm or eggs in an animal (especially if fertilisation may occur).
It recommendations include:
  • That the Home Office puts in place an expert oversight body, within the current system of animal research regulation, to oversee the most sensitive types of ACHM research.
  • Close alignment of several regulatory bodies that oversee aspects of ACHM research (particularly the Home Office and HFEA).
  • Raising international awareness of ACHM, promoting international consistency in research practice, and the development of international standards and guidance.
These recommendations should ensure that valuable and justifiable ACHM research can proceed within a robust, proportionate regulatory system, which is capable of responding to developing scientific knowledge and social attitudes, and which avoids undue bureaucracy and duplication of regulation.
The study was supported by the Department for Business, Innovation and Skills’ Sciencewise-ERC programme, the Department of Health, Medical Research Council, and Wellcome Trust. A report synopsis has been prepared by Dr Geoff Watts FMedSci.
Further information is available on The Academy of Medical Sciences website

TT2011 meeting: abstracts and awards deadlines coming soon

TT2011 Meeting: 24-26 October 2011, Trade Winds Island Grand Resort, St Pete Beach, Florida, USA
TT2011 Meeting: 24-26 October 2011, Trade Winds Island Grand Resort, St Pete Beach, Florida, USA

The 10th Transgenic Technology meeting (TT2011) will be held at the Trade Winds Island Grand Resort, in St Pete Beach, Florida, USA, on 24-26 October 2011, organized by the International Society for Transgenic Technologies (ISTT). The first deadline for submitting abstracts and applying for the various awards (Registration and Young Investigator Awards) will be due in about one month, on 30th June 2011. Accepted abstracts will be published in the scientific journal Transgenic Research, associated with the ISTT. Results from the various awards will be communicated by 16th July 2011.

Please, mark the TT2011 meeting in your agendas and don’t miss this opportunity to discuss the latest developments on Transgenic Technologies, at the International Level, with the world experts in the field. Looking froward to receive your latest work, experiments, designs, developments, your hottest research achievements using genetically modified animals.

Classical videos on transgenic and KO mice from CSHL Press available now to ISTT members

Transgenic Techniques in Mice: A Video Guide by Roger A. Pedersen, Janet Rossant, © CSHL Press, 1989
Transgenic Techniques in Mice: A Video Guide by Roger A. Pedersen, Janet Rossant, © CSHL Press, 1989

The International Society for Transgenic Technologies is most proud to announce the release of the digital versions of the two classical video guides on transgenic and knockout mice, produced by Cold Spring Harbor Laboratory Press (CSHL Press) in 1989 and 1993, that will be now available to ISTT members for learning and/or teaching purposes, from the ISTT members-only restricted area, thanks to a most generous agreement reached with CSHL Press, owner of the copyright for both videos, marketed as analogic PAL/NTSC videos.

These two videos have been, for many years, unique and the most used and helpful video reference for anyone willing to start learning/teaching in the field of genetically modified mice. The first video is “Transgenic Techniques in Mice: A Video Guide“, by Roger A. Pedersen and Janet Rossant, produced by CSHL Press in 1989. The second video is “Targeted Mutagenesis in Mice: A Video Guide“, by Roger A. Pedersen, Virginia Papaioannou, Alexandra Joyner and Janet Rossant, produced by CSHL Press in 1993.

Targeted Mutagenesis in Mice: A Video Guide, by Roger A. Pedersen, Virginia Papaioannou, Alexandra Joyner and Janet Rossant, © CSHL Press, 1993
Targeted Mutagenesis in Mice: A Video Guide, by Roger A. Pedersen, Virginia Papaioannou, Alexandra Joyner and Janet Rossant, © CSHL Press, 1993

The contents of the two videos are:

Transgenic Techniques in Mice: A Video Guide (CSHL Press, 1989)

  • Procedure 1: Dissection of oviducts. Recovery of fertilized eggs. Removal of cumulus cells.
  • Procedure 2: Recovery of 8-cell embryos. Removal of the zona pellucida. Construction of aggregation chimeras.
  • Procedure 3: Dissection of uteri. Recovery of blastocysts.
  • Procedure 4: DNA injection into pronuclei to produce transgenic embryos.
  • Procedure 5: Nuclear transfer.
  • Procedure 6: Blastocyst injection of embryonic stem cells.
  • Procedure 7: Vasectomizing male mice.
  • Procedure 8: Oviduct transfer of manipulated embryos.
  • Procedure 9: Uterine transfer of manipulated embryos.
  • Procedure 10: Recovery of 6-1/2 and 7-1/2 day embryos.
  • Procedure 11: Dissection of midgestation 12-1/2 day embryos and fetal membranes

Targeted Mutagenesis in Mice: A Video Guide (CSHL Press, 1993)

  • Demonstration 1: Starting embryonic stem (ES) cell lines
  • Demonstration 2: Making fibroblast feeder layers
  • Demonstration 3: Growing established ES cell lines
  • Demonstration 4: Targeting ES cell lines
  • Demonstration 5: Making chimeras with ES cells
  • Demonstration 6: Making a mouse

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

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.