Genetic Engineering of Human Embryos- for discussion

Commentary by Ernst-Martin Füchtbauer

The new and accelerating technical development of the CRISPR/Cas9 system opens up for the possibility of targeted genetic modifications in germline competent human embryos. This is an avenue, which until very recently has been regarded as absolutely off limits. To cross the border between genetic modifications of somatic cells and germline cells was simply not conceivable, at least in most Western countries. Indeed, the border has not yet been crossed, but we are getting closer.

In two recent papers Chinese scientists used triploid human embryos as a ‘model’ to either treat ß-thalassemia [1] or to recapitulate a spontaneous mutation in the CCR5 gene [2], which results in resistance against HIV infections. Both targets are clearly chosen due to their potential for future therapeutic application.

Shortly after the first of the two papers was published, the Board of Directors of the ISTT posted a statement [3], which among other arguments contains the following sentence:

Uses of genetic engineering in human embryos should be limited to disease mitigation for those diseases where no other option is available; we reject the idea of “designer babies”.

This raises the question whether there are at all diseases where there are no other options (now or in the future). Hereditary diseases are rarely transmitted by homozygous parents, which makes preimplantation diagnostics (PID) an obvious safer and ethically far less disputed alternative. The example, ß-thalassemia reaches in very limited populations, like the Maldives, a frequency that puts about 1% of couples at risk to be double homozygous. But still, is not a CRISPR/Cas9 based hematopoietic stem cell therapy the obvious and much easier developed therapy?

However, the case of targeting CCR5 is fundamentally different. As no one can claim that being wild type for CCR5 is a disease, this is a clear designer approach. Given that we know relatively little about the function of CCR5, one might wonder how we can be sure that it is beneficial to mutate it in a world of ever changing microbial threats. It seems that developing a CCR5 blocking drug or somatic mutations of CCR5 in HIV patients is the obvious way forward.

It is my feeling that many colleagues, some of whom I greatly admire, are beginning to accept experiments with the obvious goal to modify the human germline ‘if it is the only cure for severe diseases’. However, I have not heard one convincing example of such disease that is not in principle “treatable” or “avoidable”. Finally we should keep in mind that the Hardy-Weinberg equation ridicules all eugenic attempts to clean the population from ‘disease’ alleles.

I am increasingly concerned because the discussion in our community has, within a few months, taken an almost purely technical turn about off target risks and efficiency. We neglect many decades of thorough philosophical and ethical literature on the issue. There is more at stake than the possible treatment of a few rare diseases.

These questions are too important to just wait and see. We as ISTT members are so close to the topic that we need to have an honest and open discussion about our opinions. This blog could be a starting point and I invite/encourage you to add to this discussion.

[1] Liang, P. et al. Protein Cell (2015)

[2] Kang, X. et al. J. Assist. Reprod. Genet. (2016)


CARD – IP Mouse Sperm and Embryo Cryopreservation Course, Institut Pasteur, Paris, France, 20-24 June 2016


The International Society for Transgenic Technologies (ISTT) will proudly co-sponsor the CARD – IP Mouse Sperm and Embryo Cryopreservation Course that will be held at the renowned Pasteur Institute, in Paris, on 20-24 June 2016, organized by Naomi Nakagata (CARD-Kumamoto University, Japan, Coordinator of CARD) and Jean Jaubert (Pasteur Institute, Paris).

Recent developments from the laboratory of Prof. Naomi Nakagata (CARD-Kumamoto University, Japan) have pushed the envelope of mouse cryopreservation: i) improved female superovulation method; ii) fresh, frozen and cold storage sperm techniques; iii) optimized IVF methods. These improvements (see main references in program) have resulted in an unparalleled increase in efficiency of cryopreservation and rescue of relevant mouse lines.

The aim of this course is to introduce the newest CARD methods to researchers and technicians involved in mouse archiving and/or managing transgenic facilities and who are willing to implement these new methods in their work. These techniques will be taught directly by the team that devised them.

This course is open to anyone interested. Pre-application will be required, including, at least, a recent CV and a letter prepared by the intended participant describing how the applicant will benefit by attending this course and how relevant is the course material to his/her work. Additional documents are welcome, at the discretion of participants, including supporting letters by supervisors (where appropriate), reference letters, etc… A copy of the passport is mandatory. Applications should be submitted online, and will close on March 25th 2016.

The maximum number of participants attending this course will be 20, distributed among countries and institutions, and according the documentation provided and the interests expressed. Review and selection of participants will be done by the Teaching Committee and results will be communicated by April 15, 2016. The official language of the course will be English.

In addition to practical sessions, the course will also include several lectures of related interesting topics for the participants delivered by experts in each field.

See more at:


Screen Shot 2016-02-02 at 15.20.55

Hands-on topics:


Making pipettes and embryo handling

Superovulating/Ultra-superovulating female mice

Isolating unfertilized mouse oocytes

Isolating and cold storage/shipping of mouse cauda epididymis

Freezing/thawing mouse sperm and IVF

Fresh mouse sperm and IVF

Freezing/thawing 2-cell IVF-derived mouse embryos

Vitrification of mouse oocytes and embryos

Embryo transfer techniques in mice (oviduct, uterus, NSET)

Vasectomy of male mice (scrotal and abdominal)


Additional lectures:


The laboratory mouse origin

Historic and scientific perspectives of transgenesis methods and the future of transgenic platforms

Historic and Scientific perspectives of embryo and sperm cryopreservation

Comparing current embryo and sperm cryopreservation methods

Vitrification of oocytes and their use for IVF

Cold storage and transport of germplasm

Shipping mice, refrigerated and frozen material

Managing and handling information in cryopreservation centers

CRISPR/Cas9 and the challenges in freezing these new GEM’s

NSET: non-surgical embryo transfer

Breeding, genotyping and back-ups of GEM’s





Naomi Nakagata (CARD-Kumamoto University, Japan)

Toru Takeo (CARD-Kumamoto University, Japan)

Shuuji Tsuchiyama (CARD-Kumamoto University, Japan)

Kiyoko Fukumoto (CARD-Kumamoto University, Japan)

Yukie Haruguchi (CARD-Kumamoto University, Japan)

Tomoko Kondo (CARD-Kumamoto University, Japan)

Yumi Takeshita (CARD-Kumamoto University, Japan)

Yuko Nakamuta (CARD-Kumamoto University, Japan)

Tomoko Umeno (CARD-Kumamoto University, Japan)

Hidetaka Yoshimoto (CARD-Kumamoto University, Japan)

Ayumi Mukunoki (CARD-Kumamoto University, Japan)

Mari Iwamoto (CARD-Kumamoto University, Japan)

Fumi Takahashi (CARD-Kumamoto University, Japan)

Kristy Kinchen (Gainesville, FL, USA)

Jean Jaubert (Institut Pasteur, France)

Franck Bourgade (Institut Pasteur, France)

Angélique Vincent (Institut Pasteur, France)

Claire Lecestre (Institut Pasteur, France)

Jorge Sztein (Barcelona, Spain)

Lluís Montoliu (CNB-CSIC, Madrid, Spain)

Barbara Stone (ParaTechs, Lexington KY, USA)


Additional lectures:


Naomi Nakagata (CARD-Kumamoto University, Japan)

Toru Takeo (CARD-Kumamoto University, Japan)

Shuuji Tsuchiyama (CARD-Kumamoto University, Japan)

Jorge Sztein (Barcelona, Spain)

Lluís Montoliu (CNB-CSIC, Madrid, Spain)

Fernando Benavides (MD Anderson, Smithville, USA)

Francina Langa Vives (Institut Pasteur, Paris, France)

Michel Cohen-Tannoudji (Institut Pasteur, Paris, France)

Xavier Montagutelli (Institut Pasteur, Paris, France)

Jean Jaubert (Institut Pasteur, Paris, France)

Barbara Stone (ParaTechs, Lexington KY, USA)


For any further information contact:


Tags: Cryopreservation course, embryo cryopreservation, ISTT co-sponsorisation, IVF, practical course, sperm cryopreservation, superovulation, vasectomy, vitrification

The ISTT Calendar for year 2016

Download a free copy of the 2016 ISTT Calendar
Download a free copy of the 2016 ISTT Calendar

Happy New Year to all ISTT Members!. Here, you can download a free copy of the 2016 ISTT Calendar. This edition has been nicely prepared by ISTT Board Member Karen Brennan (Sydney, Australia), using numerous beautiful images generously provided by ISTT members. Download it, print it and use it!. Enjoy it!.


ISTT Board of Directors


June 10, 2015

Genetic engineering in animals is a process that has engendered great excitement as well as great anxiety. The technology is used to study developmental processes (using small animals such as the mouse, zebra fish, fruit fly, worm, etc.), determine gene function, and mimic human and animal disease processes. Perhaps the greatest promises of this technology are to develop and test drugs and to perform gene therapy, both of which are intended to prevent or cure disease.

Until recently, a variety of limitations made the technology impractical for all but a few species of animals (primarily mice). However, with the advent of new gene-editing systems, where components are inexpensive, readily generated in the laboratory, and applicable to virtually any species, it is now feasible to perform genetic engineering in the human embryo. Changes made in an embryo brought to term would no longer be confined to that individual, but could be passed through the germline to affect future generations.

A recent publication [Liang, P. et al. Protein Cell (2015)] brought this reality squarely into the public consciousness. In this study, the CRISPR/Cas9 system was used to edit the genome of human embryos. To their credit, the authors were careful to use only non-viable embryos. Furthermore, their detailed examination of the engineered embryos revealed both the intended and unintended modifications that resulted. This study clearly demonstrates that the CRISPR/Cas9 system is currently too imprecise and inefficient for genetic engineering of human embryos for implantation, gestation and birth.

Members of the ISTT use CRISPR/Cas9 technology, as well as other gene-editing technologies, routinely. Many of our members have had integral roles in the development of these technologies and therefore recognize the power of these systems. It is with that knowledge and foresight that the ISTT Board of Directors issues this statement (while understanding that more nuanced discussions and decisions will be needed as the technology improves):

  • Genetic engineering technology, in its current state, is error-prone and must not be used in human embryos intended for implantation.
  • Studies to test new genetic engineering technology in human embryos should be postponed until proven completely safe and effective in other species.
  • New methods of genetic engineering must be carefully assessed to ensure that risk to the human population is negligible.
  • Uses of genetic engineering in human embryos should be limited to disease mitigation for those diseases where no other option is available; we reject the idea of “designer babies.”
  • We strongly urge worldwide agreement on minimum standards for gene editing experiments in human embryos, and will promote such measures with our members. Until such standards have been established, we remain opposed to making any genetic alterations in human embryos that could be inherited by future generations.


Advances in the Generation of Genetically Modified Animal Models: International Course & Symposium, Institut Pasteur de Montevideo (Uruguay), 7-18 September 2015

Advances in the Generation of Genetically Modified Animal Models: International Course & Symposium, Institut Pasteur de Montevideo (Uruguay), 7-18 September 2015
Advances in the Generation of Genetically Modified Animal Models: International Course & Symposium, Institut Pasteur de Montevideo (Uruguay), 7-18 September 2015

The International Society for Trangenic Technologies (ISTT) proudly co-sponsors the International Course & Symposium on Advances in the Generation of Genetically Modified Animal Models, to be held at the Institut Pasteur de Montevideo (Uruguay), organized by ISTT Members Martina Crispo (Unidad de Animales Transgénicos y Experimentación, UATE, Institut Pasteur de Montevideo) and Alejo Menchaca (Instituto de Reproducción Animal de Uruguay, IRAUy), on 8-15 September 2015.

The aim is to offer a training course of excellence for researchers and technicians working in animal transgenic field. The topics will be focused on both the basic knowledge and the latest advances in transgenic technologies. The course consists of a 1st week of lectures sessions and a 2nd week of practical sessions. In addition, a mini symposium (11-12 September) is organized in order to extend the impact of the presence of the professors to other researchers, technicians and posgraduate students. Current programs for the COURSE and MINI-SYMPOSIUM.

Confirmed speakers attending this Course and mini-Symposium include:

  • Michel Cohen-Tannoudji, IPParis, France
  • Francina Langa, IP Paris, France, ISTT member
  • Ignacio Anegón, INSERM, Nantes, France, ISTT member
  • Lluis Montoliu, CNB, Spain, ISTT member
  • Jorge Sztein, consultant, Spain
  • Sylva Haralambous, HPI, Greece, ISTT member
  • Naomi Nakagata, CARD, Kumamoto U, Japan, ISTT member
  • Charles Long, Texas A&M University, USA
  • Daniel Salamone, Fagro, UBA, Argentina
  • Adrian Mutto, UNSM, Argentina
  • Marcelo Rubinstein, INGEBI, Argentina, ISTT member
  • Marcelo Bertolini, UNIFOR, Brazil

Local professors and instructors include:

  • Magdalena Cárdenas, IP Montevideo, Uruguay
  • Ana Paula Mulet, IP Montevideo, Uruguay
  • Geraldine Schlapp, IP Montevideo, Uruguay, ISTT member
  • María Noel Meikle, IP Montevideo, Uruguay, ISTT member
  • Gabriel Fernández, IP Montevideo, Uruguay
  • Ana Paula Arévalo, IP Montevideo, Uruguay
  • Martina Crispo, IP Montevideo, Uruguay, ISTT member
  • Pedro C. dos Santos, IRAUy, Uruguay
  • Natalibeth Barrera, IRAUy, Uruguay
  • Federico Cuadro, IRAUy, Uruguay
  • Alejo Menchaca, IRAUy, Montevideo, Uruguay, ISTT member

People interested in participating in this COURSE must send the COURSE Application Form to
A maximum of 20 students will be accepted for the COURSE taking into account personal qualifications.
There is no registration fee for the COURSE. Support for accommodation, per diem and local transportation will be provided to all participants from abroad. Travel expenses are not included.
People interested in participating in the MINI SYMPOSIUM must send the SYMPOSIUM Registration Form to
SYMPOSIUM fee is U$S 100.

Deadline for COURSE applications is June 28th
Deadline for SYMPOSIUM registrations is July 19th
For any further information contact:


CARD-RPCI Cryopreservation Course Report

CARD-RPCI Cryopreservation Course Report
CARD-RPCI Cryopreservation Course Report

During the past week, 15-19 September 2014, the CARD-RPCI Mouse Sperm and Embryo Cryopreservation Course was held at Roswell Park Cancer Institute, in Buffalo, NY, USA, organized by Naomi Nakagata (CARD-Kumamoto University, Japan), Aimee Stablewski (Roswell Park Cancer Institute, Buffalo, NY, USA) and Jan Parker-Thornburg (MD Anderson Cancer Center, Houston, TX, USA). This was the second CARD course organized overseas, outside Asia, after the first course organized in Madrid in October 2013. This new practical course in North America was co-sponsored by the International Society for Transgenic Technologies (ISTT). What follows is a brief course report prepared by Aimee Stablewski and Jan Parker-Thornburg, who deserve to be praised, along with Naomi Nakagata and his CARD team, for another most successful cryopreservation course. This course report can also be downloaded from here.

CARD-­RPCI Spermand Embryo CryopreservationWorkshop
September 15-­19, 2014
Roswell Park Cancer Institute, Buffalo NY, USA


The CARD-RPCI Sperm and Embryo Cryopreservation Workshop was recently held in Buffalo NY USA on the campus of Roswell Park Cancer Institute. Eighteen trainees, fifteen instructors and lecturers, and numerous vendors were hosted by Naomi Nakagata (Kumamoto University), Aimee Stablewski (Roswell Park Cancer Institute) and Jan Parker-Thornburg (MD Anderson Cancer Center) from September 15-19, 2014. Drs. Naomi Nakagata and Toru Takeo brought their team from CARD to assist the trainees in learning the latest techniques in embryo vitrification, sperm cryopreservation, in vitro fertilization, and the new method of vitrifying oocytes. In addition, Jorge Sztein instructed the trainees in methods of ovary freezing and subsequent implant after thaw; Lluis Montoliu provided lectures in current methods of embryo cryopreservation and CRISPR/Cas9 methods of generating genetically engineered mice, and Barbara Stone provided training in using the NSET method of embryo implant.

This was the first time that the CARD team provided training in North America, and the team (supported by Kumamoto University and Kyudo Co., LTD.) delivered a truly memorable experience for the trainees. Hands-on sessions were intense and intended to provide trainees with the knowledge and experience of performing the exacting CARD techniques. The eighteen trainees were composed of fourteen ISTT members and four non-members. Of these, three were from Europe, one from Australia, one from New Zealand, and the remainder from all parts of North America.

Hands on training included isolating unfertilized mouse oocytes, isolating and cold storage/shipping of mouse cauda epididymis, freezing/thawing mouse sperm and IVF using CARD frozen sperm and legacy sperm, fresh mouse sperm and IVF, cold stored sperm and IVF, freezing/thawing 2-cell IVF-derived mouse embryos, vitrification of mouse oocytes and embryos, IVF of vitrified mouse oocytes, ovary transplantation/ovary freezing, and embryo transfer techniques in mice (oviduct, uterus via NSET).

Didactic lectures were given on the topics of historic and scientific perspectives of embryo and sperm cryopreservation (by Jorge Sztein), comparing current embryo and sperm cryopreservation methods (by Lluis Montoliu), vitrification of oocytes and their use for IVF (by Naomi Nakagata), new US guidelines for the use of animals in research/IACUC (by Sandra Sexton), CRISPR/Cas9 and gene editing endonucleases (by Lluis Montoliu), development of database for managing mouse banking system (by Shuuji Tsuchiyama), shipping mice, frozen or refrigerated embryos/sperm across the world (by Toru Takeo) and freezing and transplantation of ovaries (by Jorge Sztein).

On Wednesday of the workshop, all of the participants were treated to a trip to Niagara Falls followed by an exceptional gala dinner (arranged by Aimee Stablewski) at a local winery. This short respite enabled the participants to replenish their energies for subsequent long days of IVF, vitrifications and surgery.

In all, it proved to be an exceptional workshop, with all participants achieving exceptional results in most, if not all, of the practicals. In fact, the depth of appreciation for being taught these methods became clear both during the closing session, where all instructors, vendors, and participants thanked Dr. Nakagata with a standing ovation, and immediately afterward, where many compliments were given. Perhaps, one of the participants said it best in an e-mail immediately following the course:

a big thank you to Prof. Nakagata, Dr. Toru Takeo and the entire CARD team. The way they have organised is unbelievable. As we all know in biological practical experiments, how much ever we take care, they never go according to plan and there is always a blooper. I must confess, I had my doubts that we would ever stick to the time table. I am glad that I was completely proven wrong. They have given attention to every bit of details and must have put lot of rehearsals behind this. They were fantastic. They answered all the questions and made sure everyone understood plus followed it up with the practicals. The Entire Card team was simply amazing and no other way to express my gratitude than simply saying “I bow to the entire CARD team”.”
Prasanna Kallingappa
University of Aukland
New Zealand

Aimee Stablewski and Jan Parker-Thornburg, as hosts, would like to acknowledge not only our lecturers, but also the CARD team who assisted the students, including Shuuji Tsuchiyama, Kiyoko Fukumoto, Yukie Haruguchi, Tomoko Kondo, Yumi Takeshita, Yuko Nakamuta, Tomoko Umeno, (all from Kumamoto University and Kyudo Co. LTD); and the CARD “adoptees”—Kristy (Kinchen) Williams (University of Florida) and Amar Dasari (Taconic). In addition, technical assistance was ably provided by Dawn Barnas (ISTT), Karstin Webber, Sandra Sexton and Leslie Curtin (all RPCI), who treated over 600 mice with hormones!

The workshop was extremely fortunate to have extremely generous vendor support, including our platinum sponsors: Charles River Laboratories (who supplied all of the mice used in the course), Leica Microsystems (who provided all of the microscopes used for the course as well as onsite support by Louise Bertrand), and Kyudo Co., LTD who provided support for the CARD instructors as well as onsite support by Nobuyuki Mikoda; our gold sponsors: the ISTT (who sponsored Lluis Montoliu’s participation and contributed to the gala dinner), Taconic (who provided instructional support from John Couse and Amar Dasari), and Regeneron (who sponsored a lecture by Lluis Montoliu); our silver sponsors: Hamilton Thorne (who provided instruction in sperm analysis and laser-assisted IVF by Nancy Mutch), IDEXX Bioresearch for course support, Paratechs for sponsoring Dr. Barbara Stone’s participation, Cell Preservation Solutions who provided course support and Lifor media for cold-storage, CosmoBio who provided course support, Transposagen who provided course support, and our bronze sponsors: Millipore EMD and mofa for equipment, Tokai-Hit for equipment, Eppendorf (Mike Bady) for course support and equipment, and VWR and Sarstedt for course support.

Finally, we thank our participants for their hard work and dedication to bring the newest transgenic technologies back to their institutions.

Aimee Stablewski, Roswell Park Cancer Institute, Buffalo, NY, USA
Jan Parker-Thornburg, MD Anderson Cancer Center, Houston, TX, USA

Mouse Genetics. Methods and Protocols (2014)

Mouse Genetics. Methods and Protocols (2014)
Mouse Genetics. Methods and Protocols (2014)

This is yet another interesting book in our field that has been published this year, 2014. This manual, entitled “Mouse Genetics. Methods and Protocols“, edited by Shree Ram Singh and Vinzenzo Coppola, in association with the Publisher, Humana Press/Springer, contains a collection of useful protocols covering most of the methods that can be currently applied for the genetic modification of the mouse genome. According to its presentation at the Springer web page, this book “provides selected mouse genetic techniques and their application in modeling varieties of human diseases. The chapters are mainly focused on the generation of different transgenic mice to accomplish the manipulation of genes of interest, tracing cell lineages, and modeling human diseases. (…) Each chapter contains a brief introduction, a list of necessary materials, systematic, readily reproducible methods, and a notes section, which shares tips on troubleshooting in order to avoid known pitfalls.

The table of contents of this book illustrates the variety of highly sophysticated methods, beyond standard techniques, that are discussed here in detail, around mouse genetics, including: pronuclear injection-based targeted transgenesis through Cre-loxP specific recombination, the use of recombinase-mediated cassette exchange (RMCE) strategies, several approaches for preparing and analyzing conditional mutant alleles using tamoxifen-dependent Cre recombinases, the use of ICSI for the generation of transgenic mice, the use of BACs, mosaic analysis with double markers (MADM) in mice, transposon-mediated transgenesis, overexpression of microRNAs using Rosa26-mediated recombination, the isolation of various somatic and pluripotent cells, the generation of transgenic mice through spermatogonial stem cells in vivo, and, several illustrative examples of how different mouse engineered animal models are best suited to study a variety of human diseases. Hence, this book is also complementary to other recently published manuals, since it contains a careful detailed description of new methods that are not been covered in other similar titles in the field.

The editors of this book, Shree Ram Singh and Vinzenzo Coppola, have counted with the generous expertise shared and provided by a very large list of co-authors, including some ISTT members: Masato Ohtsuka, Kazuhito Sakamoto, Channabasavaiah B. Gurumurthy, Kay-Uwe Wagner, Petra Kraus, V. Sivakamasundari, Xing Xing, Thomas Lufkin, Anton J.M. Roebroek, Bart Van Gool, Kun-Hsiung Lee, Susanne Feil, Jana Krauss, Martin Thunemann, Robert Feil, Pedro N. Moreira, Lluis Montoliu, Jane Beil, Thorsten Buch, Sheng Ding, Tian Xu, Xiaohui Wu, Hui Zong, Claudia Piovan, Foued Amari, Francesca Lovat, Qun Chen, Olga Simmons, Esther M. Bolanis, Jian Wang, Simon J. Conway, Kanika Jain, Paul J. Verma, Jun Liu, Pollyanna Agnes Goh, Michael D. Williams, Wilson Wong, Amanda Rixon, Sarang N. Satoor, Anandwardhan A. Hardikar, Mugdha V. Joglekar, Andrei M. Vacaru, Joseph Vitale, Johnathan Nieves, Margaret H. Baron, Kristbjorn Orri Gudmundsson, Kevin Oakley, Yufen Han, Yang Du, Lalit Sehgal, Abul Usmani, Sorab N. Dalal, Subeer S. Majumdar, Spencer W. Luebben, Naoko Shima, Tsuyoshi Kawabata, Robert M. Hoffman, Viive M. Howell, Emily K. Colvin, Vishalakshi Chavali, Shyam Sundar Nandi, Paras Kumar Mishra, Julia Lorenz, Susanne Grässel, Ganesan Ramesh, Punithavathi Ranganathan, Santhakumar Manicassamy, Indumathi Manoharan, Deepak P. Patil, Holly D. Kristensen and Yogesh Shouche.

This new book will be added to the collection of Springer books published on animal transgenesis and animal genetics for which ISTT members are entitled to a 33% discount, as one of the many benefits associated with the ISTT membership.



Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)

Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)
Transgenic Animal Technology. A Laboratory Handbook (3rd edition, 2014)

Twenty years after the publication of the first edition and twelve years after the release of the second edition of this book, Carl A. Pinkert (Auburn University, College of Veterinary Medicine, Auburn, AL, USA) in association with Elsevier, releases now the third edition of his famous transgenic manual: “Transgenic Animal Technology. A Laboratory Handbook. 3rd edition, 2014“. As it will be familiar to readers of the two previous editions of this useful and unique handbook, this is not only a manual to understand how to make a transgenic mouse. This handbook looks beyond mice and it contains protocols to prepare a wide variety of genetically-modified animals, including: rats, rabbits, poultry, fish, pigs, ruminants and non-human primates. In addition, this compilation of helpful methods includes a number of chapters devoted to understand and improve all steps of transgenesis, from vector design, analysis of transgene integration and the evaluation of transgene expression. Finally, the book also includes cryopreservation methods, an embryo culture section, a review of standard nomenclature and a selection of databases and internet resources currently available in the field.

This handbook is a worth addition to any library, laboratory or transgenic facility, complementary to other available manuals on the subject, but unique in the sense that it exquisitely illustrates current transgenic methods that can be applied to a wide variety of animal species, beyond mice.

Carl A. Pinkert has been helped in his outstanding Editorial task by a large group of co-authors, experts in their subjects, including some ISTT members: Satoshi Akagi, Anna V. Anagnostopoulos, Benjamin P. Beaton, Cory F. Brayton, Steve Brown, Anthony W.S. Chan, Tom Doetschman, Rex A. Dunham, David A. Dunn, Janan T. Eppig, Almudena Fernandez, Tatiana Flisikowska, Vasiliy Galat, Robert A. Godke, Philip Iannaccone, Michael H. Irwin, Larry W. Johnson, Yoko Kato, Teoan Kim, Alexander Kind, Bon Chul Koo, Mo Sun Kwon, Daniel J. Ledbetter, Michael J. Martin, Kazutsugu Matsukawa, Colin McKerlie, Lluis Montoliu, Paul E. Mozdziak, Akira Onishi, Paul A. Overbeek, James N. Petitte, L. Philip Sanford, Jorge A. Piedrahita, Wendy K. Pogozelski, H. Greg Polites, Edmund B. Rucker III, Marina Sansinena, Angelika Schnieke, Kumiko Takeda, James A. Thomson, Ian A. Trounce, Yukio Tsunoda, Cristina Vicente-Garcia, Kevin D. Wells, Richard N. Winn and Curtis R. Youngs.

TT2014 abstracts: submission deadline is approaching (30 June)

TT2014 abstracts: submission deadline is approaching (30 June)
TT2014 abstracts: submission deadline is approaching (30 June)

From the International Society for Transgenic Technologies (ISTT) we warmly invite and encourage you all to submit your most recent and exciting results and developments in animal transgenesis to be presented at the forthcoming 12th Transgenic Technology (TT2014) meeting, which will be held in Edinburgh, Scotland, UK, on 6-8 October 2014. Deadline for submitting abstracts for the TT2014 meeting is June 30.
To submit an abstract please visit this TT2014 meeting web page.

All TT2014 participants are encouraged to submit their work as an abstract for poster presentation at the TT2014 meeting. Authors are requested to submit an abstract with the following requirements:

  • Title (max. 25 words)
  • Name authors and affiliations (first author is the presenting author).
  • Text of the communication (max. 400 words).
  • Abstracts should be submitted no later than June 30, 2014.

Accepted abstracts will be published in the scientific journal Transgenic Research (Springer), to which the ISTT is associated.

Posters will be on display in the exhibition area throughout the duration of the meeting. Poster boards are 1.00m wide x 2.00m high and we recommend posters do not exceed 1.50m in length. A supply of Velcro tabs will be available at the venue. No screws or double-sided adhesive tape will be allowed due to the damage they can cause to the boards.

Best Poster Awards
All posters presented at the TT2014 meeting will be eligible for one of the ISTT Best Poster Awards, generously sponsored by Charles River, Inc.

Oral Presentations
A limited number of abstract submissions will be selected and invited to present their findings in the form of a short oral presentation within the main meeting program. Should you be interested in being considered to speak at the meeting please select the appropriate option when submitting your abstract.

Abstracts are invited on all aspects of Transgenic Technologies, including the conference themes as listed below:

  • New technologies in animal transgenesis
  • Embryo stem cells
  • Target nucleases or Editing nucleases (ZFNs, TALENs, CRISPRs)
  • Large-scale phenotyping
  • Animal Biotechnology
  • Imaging with transgenic animals
  • Mouse models of human disease
  • Zebrafish models of human disease and transgenesis
  • Animal ethics and welfare

We are looking forward to receiving your exciting works to discuss the latest development on animal transgenesis!. See you all in Edinburgh!

Meeting report: Promoting the international exchange of mouse mutant resources

Infrafrontier-IMPC workshop: Promoting the international exchange of mouse mutant resources, Munich, Germany, 8-9 May 2014
Infrafrontier-IMPC workshop: Promoting the international exchange of mouse mutant resources, Munich, Germany, 8-9 May 2014

This is a brief meeting report on the INFRAFRONTIER /IMPC workshop: Promoting the international exchange of mouse mutant resources, which was held in Munich, Germany, on 08-09 May 2014.
As indicated in the corresponding Infrafrontier web page: “The main objectives of the workshop were to discuss how to simplify the international exchange of mouse mutant resources and to define the procedural changes to achieve it, to review the key issues facing the mouse community and mouse repositories as well as focus on IP issues and to present best practices in sharing research tools. The workshop was targeted at the directors of major mouse repositories, IP and technology transfer experts, representatives of scientific journals and funders and attracted the attention of 70 participants.” Delegates from major mouse repositories (JAX, MMRRC, EMMA, CMMR, RIKEN BRC, CARD, MARC), mouse international projects and consortia (EUCOMM, EUCOMMTOOLS, KOMP, KOMP2, IKMC, IMPC, KMPC), other related consortia (SGC), scientific journals (Nature, PLOS), funding agencies (NIH), companies (BioDoc, Charles River, AddGene), associations (AMMRA, AMPC, FELASA, EARA), TTOs and lawyers from numerous institutions and end-users gathered to discuss about how to best promote the international exchange of mouse mutant resources.

This workshop was funded by the EC FP7 InfraCoMP project. InfraCoMP’s main objective is to coordinate the collaborative efforts between the Infrafrontier Research Infrastructure and the International Mouse Phenotyping Consortium (IMPC). The scope of this Infrafrontier-IMPC workshop in Munich included various major topics, such as:

  • to discuss about simplified procedures to effectively exchange mouse mutant resources among repositories and between repositories and end-users/customers, trying to review and fix all restrictions preventing from adequately sharing major mouse mutant resources.
  • to review the key issues currently faced by the mouse community and mouse repositories, including emerging new genome editing technologies (ZFNs, TALENs, CRISPRs) and the role of mouse archives in the international exchange of mouse mutant resources
  • to discuss on IP issues and the administrative paperwork usually associated with any transactional international negotiation involving licenses and MTAs
  • to showcase best practices, examples of successful sharing research tools that could be applied on sharing mouse mutant resources

This workshop represented a continuation towards the eventual application of the agreements included in the so-called Rome Agenda, published in 2009 (Schofield et al. 2009, Nature) where the major headlines, best practices and recommendations concerning the deposit and sharing of biological resources, including mice, ES cells and germplasm, under the least restrictive terms possible, had been already discussed and identified but, unfortunately, not sufficiently widespread nor systematically followed, in spite of new initiatives adopted by some funding agencies, enforcing public-access policies for materials associated with projects funded by the NIH or the Wellcome Trust in order to receive the allocated funds.

The impact of the new genome editing technologies on current mouse consortia and mouse archives was discussed at length and in depth, from various angles and by different speakers. It is obvious that a new logic has emerged, the updated mouse genetics toolbox and its widespread among scientists enables them to generate their mouse mutants of interest through alternative, often faster approaches. Instead of considering the new endonuclease-mediated mutations solely a threat for traditional approaches, based on ES cell clones (however using higher genetic and quality-controlled standards), it was finally interpreted as an opportunity for mouse consortia and repositories. For example, the easier and faster generation of new mouse mutations could help finishing the functional annotations of the mouse genome, for all these loci that could not be targeted or, if targeted, did not result in the corresponding mouse strain through IKMC-IMPC current approaches.

The description of innovative shipment methods, for refrigerated biological materials, or using dry-ice, as compared to the standard but more complex liquid-nitrogen dry shippers was also discussed in order to make the distribution of mouse mutant resources cheaper and easier. The new set of sperm and oocyte cryopreservation methods and the optimized associated IVF procedures, as reported by CARD, Kumamoto University, in Japan, have also greatly contributed to promote the international exchange of mouse mutant resources, avoiding the always difficult and expensive shipment of live research laboratory animals.

The legal agreements, such as Material Transfer Agreements (MTAs), governing the access to mouse mutant resources were also discussed extensively. The complexity of some of these MTAs and the often long administrative process involved for executing them, unnecessarily extends the time required to access to a given mouse mutant strain deposited in a major repository for academic use. Interesting analyses of common practices observed within the international mouse community and applied by mouse consortia were presented (Bubela et al. 2012; Mishra and Bubela, 2014). The overall recommendation was, whenever possible, avoid using specific MTAs and favor the unrestrictive distribution of mouse resources through simpler “conditions of use”, as regularly applied by The Jackson Laboratory (JAX) to all their mouse strains, and by EMMA-INFRAFRONTIER, for mouse lines non-associated to specific MTAs, in order also to reduce the administrative time to the minimum. In case MTAs should be included, for academic non-commercial use, the recommendations discussed were to simplify, and unify, the document as much as possible, ideally without requesting to disclose the field of use, without imposing reach through on modifications of the received materials and clearly defining third-party use after permission has been obtained. Attribution should also be clearly encouraged. Examples of simplified MTAs, also including useful institutional versions of these agreements, can be found at KOMP. The model deployed by AddGene, a non-profit organization dedicated to efficiently distribute plasmids among the scientific community, using also simple MTA procedures, was also presented as an example of successful solution.

Overall, this intense 2-day Infrafrontier-IMPC workshop fulfilled its aims and expectations. All stakeholders in the field could openly express their opinions, fears, opportunities, problems and solutions. The Organizers should be praised for their selection of speakers, topics and participants. Now it will be the time for the most difficult part: converting the agreements and recommendations into realities, while ensuring that researchers in academia, using mouse mutant resources, have an easier, simpler and faster access to mice and/or their associated products, for the benefit of science, and knowledge advance.

Infrafrontier-IMPC workshop: Promoting the international exchange of mouse mutant resources, Munich, Germany, 8-9 May 2014
Infrafrontier-IMPC workshop: Promoting the international exchange of mouse mutant resources, Munich, Germany, 8-9 May 2014