11th ISTT Prize Awardee – Dr. Mario Capecchi

Dr. Mario Capecchi
Dr. Mario Capecchi

 

Houston TX, USA. 26 October, 2016.

The International Society for Transgenic Technologies (ISTT, Inc.) is delighted to announce that the 11th ISTT Prize will be awarded to Dr. Mario Capecchi for his seminal work on homologous recombination in embryonic stem cells, for which he was awarded the Nobel Prize in 2007. The ISTT Prize is awarded to investigators who have made outstanding contributions to the field of transgenic technologies. The selection of the 11th ISTT Prize winner, Dr. Capecchi, was made by the ISTT Prize Committee [composed of the ISTT President (Jan Parker-Thornburg), ISTT Vice-president (Benoit Kanzler), and the CEO of genOway (Alexandre Fraichard) (the company that generously sponsors the award), and all previous ISTT prize awardees].

The committee unanimously agreed that Dr. Capecchi’s work was essential for the field of transgenesis, having opened up the field to the possibility of generating exact genetic mutations in the mouse genome. At the time when his discoveries were published, the ability to generate transgenic animals by pronuclear injection had recently been published, and was rapidly becoming an essential method to flesh out how genes would both be regulated, and would function. However, while standard transgenesis could answer many genetic questions, it was still limited, as the gene being interrogated was still intact. To truly make leaps forward, it was essential to specifically mutate that gene, either by removing it (to generate a gene knock-out) or to mutate it (to recapitulate a genetic mutation by knock-in). With the culture and injection of ES cells described by Dr. Martin Evans, it became apparent to Drs. Smithies and Capecchi (all three being co-recipients of the 2007 Nobel prize) that such cells could be used for the introduction of mutations into the genome. Homologous recombination of an exogenous DNA into the exact gene to be replaced can occur in very rare circumstances. In his 1986 Cell publication, Dr. Capecchi showed that 1 in 103 cells in culture would undergo the process using homologous recombination. The following year, Capecchi successfully knocked out the Hprt gene in mouse ES cells and Smithies independently demonstrated repair of the gene. Dr. Capecchi gave the process the name by which we know it today, “gene targeting”. A critical aspect of gene targeting (and one that is still essential today) was that both positive and negative selection processes could be used to identify those cells that had undergone homologous recombination, a process published in 1988 in Nature by Dr. Capecchi.

Dr. Capecchi was born in Verona, Italy in 1937, the only child of an Italian father (lost in WWII) and an American mother (detained in and later released from a Nazi concentration camp). He immigrated to the USA in 1946 with his mother to live in Pennsylvania. After graduation from the George School, he attended Antioch College in Yellow Springs Ohio, USA where he majored in chemistry and physics. He began his graduate work at MIT in physics and mathematics, but after becoming interested in molecular biology, moved to Harvard University to study with James Watson, and changed the course of his career. Dr. Capecchi was on the faculty at Harvard until 1973, when he moved his laboratory to the University of Utah. His seminal work describing gene targeting was performed at that institution. During his extremely productive career, some of the awards that Dr. Capecchi has been given include the Kyoto Prize in Basic Sciences (1996), The Premio Phoenix-Anni Verdi for Genetic Research Award, Italy (2000), the 33rd Jiménez-Diaz Prize for Contributions to Medical Genetics, Spain (2001), the Albert Lasker Award for Basic Medical Research (2001), and the American Association of Cancer Research Lifetime Achievement Award (2015). He has been elected to the European Academy of Sciences (2002), the American Academy of Arts and Sciences (2009), the National Academy of Medicine (2015) and has been given honorary doctorate degrees from institutions in Italy, the UK, and Israel. Dr. Capecchi remains an active researcher, with seven research publications in 2015, and six current 2016 publications either printed, submitted, or in press.

We are most pleased that Dr. Capecchi has agreed to receive the ISTT Prize to be given at TT2017, thus joining the list of previously honored scientists, including Janet Rossant (2014), Allan Bradley (2013), Ralph Brinster (2011), Francis Stewart (2010), Brigid Hogan (2008), Charles Babinet (2007), Andras Nagy (2005), Qi Zhou (2004), Kenneth McCreath (2002) and Teruhiko Wakayama (2001). All ISTT Prize winners are given an honorary ISTT membership and a unique silver sculpture representing a mouse blastocyst created by the Hungarian artist Béla Rozsnyay. Dr. Capecchi will receive his prize at the 14th Transgenic Technology Meeting (TT2017) that will be held at the Snowbird Resort outside of Salt Lake City, Utah USA on 1-4 October, 2017.

Selected References from Dr. Capecchi’s lifetime achievements:

Folger, K. R., K. R. Thomas and M. R. Capecchi (1984). Analysis of homologous recombination in cultured mammalian cells. Cold Spring Harbor Symp. Quant. Biol. 49:123-138. PMID: 6099232.

Thomas, K. R., K. R. Folger and M. R. Capecchi (1986). High frequency targeting of genes to specific sites in the mammalian genome. Cell 44:419-428. PMID: 3002636.

Wong, E. A. and M. R. Capecchi (1986). Analysis of homologous recombination in cultured mammalian cells in a transient expression and a stable transformation assay. Somat. Cell Mol. Genet. 12:63-72. PMID: 3003931.

Thomas, K. R., and M. R. Capecchi (1986). Introduction of homologous DNA sequences into mammalian cells induces mutations in the cognate gene. Nature 324:34-38. PMID: 3785372.

Thomas, K. R. and M. R. Capecchi (1986). Targeting of genes to specific sites in the mammalian genome. Cold Spring Harbor Symp. Quant. Biol. 51:1101-1113. PMID: 3472755.

Wong, E. A. and M. R. Capecchi (1987). Homologous recombination between coinjected DNA sequences peaks in early to mid-S phase. Mol. Cell. Biol. 7:2294-2295. PMID: 3600663.

Thomas, K. R. and M. R. Capecchi (1987). Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51:503-512. PMID: 2822260.

Mansour, S. L., K. R. Thomas and M. R. Capecchi (1988). Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: A general strategy for targeting mutations to nonselectable genes. Nature 336:348-352. PMID: 3194019.

Capecchi, M. R. (1989). Altering the genome by homologous recombination. Science 244:1288-1292. PMID: 2660260.

Thomas, K. R., C. Deng and M. R. Capecchi (1992). High-fidelity gene targeting in embryonic stem cells by using sequence replacement vectors. Mol. Cell. Biol. 12:2919-2923. PMID: 1620105.

Deng, C. and M. R. Capecchi (1992). Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Mol. Cell. Biol. 12:3365-3371. PMID: 1321331.

Capecchi, M. R. (1995). A personal view of gene targeting. In Accomplishments in Cancer Research 1994. (J. G. Fortner and J. E. Rhoads, Ed.) Philadelphia: J. B. Lippincott, pp. 67-78.

Capecchi, M. R. (2000). How close are we to implementing gene targeting in animals other than the mouse? Proc. Natl. Acad. Sci. USA 97:956-957. PMID: 10655465.

Capecchi, M.R. (2001). Generating mice with targeted mutations. Nature Med. 7:1086-1090. PMID: 11590420.

Austin, C.P., J.F. Battey, A. Bradley, M. Bucan, M.R. Capecchi, F.S. Collins, W.F. Dove, G. Duyk, S. Dymecki, J.T. Eppig, F.B. Grieder, N. Heintz, G. Hicks, T.R. Insel, A. Joyner, B.H. Koller, K.C. Lloyd, T. Magnuson, M.W. Moore, A. Nagy, J.D. Pollock, A.D. Roses, A.T. Sands, B. Seed, W.C. Skarnes, J. Snoddy, P. Soriano, D.J. Stewart, F. Stewart, B. Stillman, H. Varmus, L. Varticovski, I.M. Verma, T.F. Vogt, H. von Melchner, J. Witkowski, R.P. Woychik, W. Wurst, G.D. Yancopoulos, S.G. Young and B. Zambrowicz (2004). The knockout mouse project. Nat Genet.36(9): 921-4. PMID: 15340423.

Wu, S., G. Ying, Q. Wu, and M.R. Capecchi (2007). Towards simpler and faster genome-wide mutagenesis in mice. Nat Genet. Jul;39(7):922-30. PMID: 17572674.

Li, S., Lan, H., Men, H., Wu, Y., Li, N., Capecchi, M.R., Bryda, E.C., and S. Wu (2016). Derivation of Transgene-Free Rat Induced Pluripotent Stem Cells Approximating the Quality of Embryonic Stem Cells. Stem Cells Transl Med. Sep 13. [Epub ahead of print]

Du, X., Feng, T.,Yu, D., Wu, Y., Zou, H., Ma, S., Feng, C., Huang, Y., Ouyang, H., Hu, X., Pan, D., Li, N., Capecchi, M., and S. Wu(2015). Barriers for the Derivation of Germline Competent Porcine Pluripotent Stem Cells. Stem Cell and Development (submitted).

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) http://dx.doi.org/10.1007/s13238-015-0153-5

[2] Kang, X. et al. J. Assist. Reprod. Genet. (2016) http://link.springer.com/article/10.1007%2Fs10815-016-0710-8

[3] http://www.montonerin.es/isttlegacy/isttblog/?p=1581

TT2016 – President’s Synopsis

The opening night of TT2016 was momentous promising subsequent days filled with good friends and good science. We welcomed more than 700 delegates who attended, and then proceeded to hear a wonderful talk from Andras Nagy (2005 ISTT Prize winner). Andras’ talk was followed by a delicious buffet with wine, friends, colleagues and music. The opening proved to be an excellent portent of what was to come. Over the next three days, we heard many excellent talks—talks that encompassed the use of transgenic technologies, and especially CRISPR/Cas9 technology.

Charles Gersbach presents CRISPR/Cas9 modification of the dystrophin gene
Charles Gersbach presents CRISPR/Cas9 modification of the dystrophin gene

We heard about methods to ameliorate muscular dystrophy, to humanize large animals for xenotransplantation, to make swine resistant to an endemic disease, and to examine infertility in humans. We discussed technologies that would use epigenome to target the “regulome”, that would examine non-coding areas of the genome, that would recapitulate immune syndromes in ES cells, and that would allow us to assess phenotypic changes in embryonic lethal mutant mice using imaging. We learned both the history behind the CRISPR/Cas9 system, and newer CRISPR systems that are in development. We discussed ethics, gene drive, non-injection technologies, new injection technologies, and methods of generating many more oocytes in mice. There were seventeen abstracts chosen for full presentation, examining technological developments, large CRISPR/Cas9 initiatives, and transposon-mediated transgenesis. The remainder of the more than 125 abstract submssions were displayed throughout the meeting in the spacious poster room. Three were chosen as Poster Award winners, including Vera Jansen (Optogenetic tools to study cAMP signaling in cilia and flagella), Charles-Etienne Dumeau (Efficient method for the isolation of functional single cell from the ICM of mouse blastocyst), and Hiromi Miura (Generation of knockdown mice by CRISPR/Cas9-based targeted insertion of artificial miRNA sequence). On the last day, the ISTT Young Investigator Award (sponsored by inGenious Targeting Laboratory) was given to Pablo Ross based on his work developing ES cells in farm animals.

Charles River Representative, Iva Morse, and Jan Parker-Thornburg and Elizabeth Williams (ISTT, Inc.) present the Best Poster Awards to Vera Jansen (absent), Charlie
Iva Morse (Charles River), and Jan Parker-Thornburg and Elizabeth Williams (ISTT, Inc.) present the Best Poster Awards to Vera Jansen (absent), Charles-Etienne Dumeau, and Hiromi Miura (represented by Masato Ohtsuka).
Thomas Zeyda (inGenious Targeting Laboratory) and Jan Parker-Thornburg (ISTT President) present the Young Investigator Award to Dr. Pablo Ross, UC Davis.
Thomas Zeyda (inGenious Targeting Laboratory) and Jan Parker-Thornburg (ISTT President) present the Young Investigator Award to Dr. Pablo Ross, UC Davis.
TT2016 - Cryopreservation Workshop presentation by Lluis Montoliu.
TT2016 – Cryopreservation Workshop presentation by Lluis Montoliu.

The meeting was preceded by two workshops—one on programmable nucleases (headed by Radislav Sedlacek) and one on cryopreservation (led by Martin Fray, INFRAFRONTIER). Those who attended the workshops were very pleased with the learning opportunities that were afforded them. In addition, immediately following the meeting was one additional workshop on zebrafish transgenesis (Leads: Petr Bartunek, Zbynek Kozmik, Christian Mosimann and Graham Lieschke). All of the workshops were well-attended and greatly appreciated!

First Orbis pictus lecture given by Richard Behringer.
First Orbis pictus lecture given by Richard Behringer.

There were a number of new initiatives at TT2016. We had Orbis pictus lectures—lectures designed to use pictures and clear descriptions to demonstrate answers to a problem. Richard Behringer gave an excellent, encyclopedic presentation of methods of producing genetically modified animals in a vast variety of species. Later, Thomas Boehm described how lymphoid organs developed throughout evolution to the point where vertebrates now have a thymus. Also, for the first time, we had concurrent sessions. Delegates needed to choose whether to hear about ethics in animal use, or new injection and superovulation technologies. Overall, the scientific program was exceptional!

Departing Board members
Presentation of thank-you gifts to departing ISTT Board members – Wojtek Auerbach (absent), Boris Jerchow, and Tom Fielder.

The ISTT, Inc. held its third General Assembly just prior to the Gala Dinner. During that meeting, we sadly said goodbye to three departing Board members: Tom Fielder, Boris Jerchow and Wojtek Auerbach. We also reviewed ISTT finances, membership, committee activities, and interactions with our affiliated organizations. One new ISTT initiative that was presented was an outreach committee to our members (and non-members) who perform transgenic technologies in non-rodent (generally large-animal) species. The ISTT large animal group will be headed by Martina Crispo and Bruce Whitelaw. The meeting ended with a presentation inviting membership to attend TT2017 in Salt Lake City, Utah, USA, hosted by Susan Tamowski.

A wonderful time at the Zofin Palace.
A wonderful time at the Zofin Palace.

The social program prepared by our Czech colleagues was also amazing. Delegates enjoyed the opening buffet with traditional Czech music. However, it was the Gala Dinner that proved to be the high point of the social program. The Zofin Palace was full with partygoers. The wine flowed freely, the food was wonderful, and the string quartet (plus clarinet) fantastic as well. Overall, TT2016 can be considered as one of the best TT meetings ever, and I am proud, as ISTT President, that we helped to host such a wonderful meeting. Thanks so very much to the organizers—Radislav Sedlacek, Inken Beck and Nicole Chambers. Due to their amazing work, the ISTT has again had a successful TT meeting!

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: http://www.pasteur.fr/fr/enseignement/ateliers/mouse-sperm-and-embryo-cryopreservation-course#sthash.BKcGh9VV.dpuf

 

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

CARD

 

Instructors:

 

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: enseignement@pasteur.fr

 

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

Report on the 2nd Oceania Transgenic Technology/Cryopreservation Symposium

2nd oceania symposium

The 2nd Oceania Transgenic Technology and Cryopreservation Symposium was held at the University of Tasmania’s Medical Sciences Precinct, Hobart, Australia on the 18th-19th of November, 2015. The meeting was a great success with for 48 participants from 23 research institutions across Australia, New Zealand, Japan and the USA and with the support of 9 sponsoring companies. The Organising Committee comprised of: Paul Scowen-chair and host (UTAS), Elizabeth Williams (University of Queensland Biological Resources), Kevin Taylor (Australian BioResources,) Irma Villaflor (Children’s Medical Research Institute, Westmead), Tanya Templeton (Australian Phenomics Network, Monash) and Karen Brennan (Victor Chang Cardiac Research Institute). Once again, it provided an opportunity for networking, for keeping up to date with the latest developments in transgenic technologies and for sharing knowledge through expertise, round table discussions and hands-on experience.

Highlights of the Programme:

The symposium began with a warm welcome from the host, Paul Scowen, and quickly moved on, beginning with a session of talks related to assisted reproduction techniques and their applications. Dr. Toru Takeo (Kumamoto University, Japan) , started by showing that ‘ultra-ovulation’ of female mice through the administration of a novel anti-serum (soon available as a commercial product) allows for consistent collection of a large number of usable oocytes from a single mouse. Combined with the IVF media products developed by the same group, mouse IVF techniques are now even further enhanced. Elizabeth Williams (UQBR) demonstrated that the CARD embryo vitrification technique can be adapted to use either straws or vials as the cryostorage vessel, allowing facilities to update their freezing techniques without having to change their liquid nitrogen storage equipment. An entertaining presentation was delivered by Prof. John McLaughlin, showing that cryovials can be easily converted with the new ‘Cryofork spatula’ to allow for easy vitrification of embryos in small media volumes. Rodrick Rupan described the challenges faced in rederiving immunodeficient colonies and how they were overcome. Continuing the rederivation and IVF theme, Mary-anne Migotto explained how UQBR have used IVF techniques to offer a rapid and large scale rederivation service. Tanya Templeton expanded upon her talk from the first Oceania symposium, giving an update on developments in ICSI technique and use of piezo methodology at Monash University. The session concluded with Julie Stanley’s talk on sperm cryopreservation at WEHI.

 

The second session covered various aspects of rodent health screening, biosecurity, monitoring, risk assessment and challenges when dealing with genetically modified mice models. Dr Trasti presented key concepts for a rodent sentinel program, establishing exclusion criteria, sample collection, interpretation of report and preparation of an action plan in case of an outbreak situation. Dr Villaflor discussed recommendations on which pathogens to monitor, issues with working with humanized immune deficient mice, quality control system and her experiences with adapting the latest trend in laboratory animal health monitoring programs. Dr Stevenson shared his insights on an appropriate scoring system for determining what agents and factors pose a risk to animal facilities housing genetically modified animals.

 

The third and final session of the day covered aspects of quality control. Sue Raboczyj (UQBR) and John Swift (OHIO) explained clearly the Quality Assurance and Control requirements for operating transgenic facilities with multiple clients, and ensuring the ongoing maintenance of a growing archive of cryopreserved material. Dr. Takeo also presented the methods used to establish a robust infrastructure at CARD (Kumamoto University). The day concluded with a personal account of undergoing the NATA accreditation process from Barbara Hunt (ANU).

Day 2 began with an extremely informative session on CRISPR technology. Kevin Taylor began with a talk on the introduction to CRISPR, the theory behind its evolution, the applications and advances based on his experiences at ABR in Moss Vale. Kevin also mentioned about the new refinements such as SCR7 and also briefly talked about electroporation. Fabien Delerue (UNSW) reported on the traditional methods utilised for creating knock-outs versus CRISPR/Cas9 and followed on with details of some of the projects that he is undertaking at UNSW and what he has done to refine how he carries out his projects being that he is a small outfit. Sandie Piltz’s work at the University of Adelaide is one of the pioneering Australian groups to start working with this technology, she discussed her personal experience with this methodology and what they have done to refine the technique, such as donor strains and ages, recipient strains, needle parameters, injection reagents and concentrations, environmental influences and superovulation techniques. Fiona Waters (WEHI) gave an introduction on the CRISPR/Cas9 methodology and then launched into the experiences from the WEHI team on how they have utilised and transferred their skills to this technique and the refinements they have made. Dirk Truman (APN) talked about the services that the APN provides, their efficiencies and that it is one of the first non-for-profit services offering this type of genome editing to Australian Researchers. Dirk also discussed the different species that are being used for this type of genome editing, off target effects, and what APN has done to refine the technique. The last talk for this session, Michelle Brownlee (ABR), talked about the day in the life of a microinjectionist. She discussed the strains of mice used, superovulation techniques, injection techniques, embryo transfer methods, refinements and troubleshooting.

The following session encompassed the administration and training aspects of working in a transgenic animal facility. Tracy Doan (UQBR) gave an overview of the database systems used and administrative support provided to help coordinate the cryopreservation and rederivation services at the Transgenic Animal Service of Queensland. Kevin Taylor (ABR) and Keri Smith (UTas) presented their respective training programs based on structure which also opened up lively discussions on different training opportunities offered in various institutions and suggestions to meet training needs of staff members. Dr David Steele who is Institutional Biosafety Committee (IBC) Chairman at the University of Tasmania, outlined the structure and important role and function of the IBC, including dealing with GMOs when conducting research in accordance with legislation, codes of practice and licensing requirements.

It is envisioned that future meetings will serve to strengthen our collaboration efforts with colleagues from various institutions doing the same type of work.

Newly elected 2016 ISTT Board of Directors members

ISTT BOD electees

Branko Zevnik Lynn Doglio Peter Hohenstein

The official results of the recent election for the ISTT Board of Directors are in, and we congratulate Branko Zevnik, Lynn Doglio and Peter Hohenstein on their election to the ISTT Board. Branko Zevnik (Cologne, Germany) is the head of the in vivo Research Facility at the University of Cologne. Lynn Doglio (Chicago, USA) is the Director of the Transgenic and Targeted Mutagenesis Laboratory at Northwestern University. Peter Hohenstein (Edinburgh, United Kingdom) is a Group Leader and Chair of the Small Animal Facility Management Committee at The Roslin Institute. All three candidates received the support of the majority of voting ISTT members. While they will start their three-year terms at TT2016 to be held in Prague, Czech Republic, they will be immediately appointed as Board members-elect. This will allow them to interact with the rest of the ISTT Board members and become familiar with the administration and management of the ISTT. Congratulations to all of them!

In addition, the ISTT would like to express its respect and sincere appreciation for the commitment and participation of all the candidates who were involved in the ISTT election process. As well, we would like to thank Tom Fielder, Wojtek Auerbach, and Boris Jerchow for their service on the ISTT Council and Board of Directors. These three Board members have contributed greatly to the management of the ISTT (and will continue to do so until TT2016). It is due in large part to the contributions of our members that we have the vibrant Society that we do.

TARC X Meeting Report

20150809_084915 20150817_103359 20130810_163007Tahoe City, California, USA

August 9 – 13, 2015

“What if . . . we had cows that did not have horns? We do! This is a naturally-occurring mutation, and these are called “polled” (or, hornless) cows. This is a great benefit to the cattle industry, as this reduces the amount of trauma that cows can cause each other. Unfortunately, there are only a few types of cows that contain the mutation causing the polled phenotype. Other cows must have their horns removed to safely interact with each other in groups and their handlers. You can see that this type of “surgery” could also cause animal welfare issues.

But, what if we could transfer the naturally-occurring mutation from one type of cow to another? This can be accomplished by breeding the mutation into non-polled cattle. Keeping in mind that the time for gestation in cattle is 9 months, and then the time to sexual maturity could be another one to one and a half years, the time needed to do the number of crosses to generate this mutation in a new strain of cattle could be significant—one breeder’s lifetime. But (again, another “but”), what if we could introduce this mutation in a single generation by genetic engineering and leave no footprint behind—just this ONE MUTATION. It is now possible to do this using the CRISPR/Cas9 system; one could introduce the mutation and carefully characterize the animals that result to insure that there are no additional changes in the genome—no footprints. You could argue that this would be incredibly beneficial for animal welfare issues and for the benefit of those who care for these animals.”

This is the type of discussion that can result, based on the research presented at the Tenth Transgenic Animal Research Conference (TARC X) [http://www.cevs.ucdavis.edu/confreg/?confid=732] just completed in Lake Tahoe, California, USA. The discussions and talks centered around transgenic animals other than mice, including cows, sheep, goats and pigs, as well as avians (chickens), rabbits, and even mosquitoes! An especially valuable addition to the signature 10th Conference was the inclusion of reviews of different aspects of the technology given at the start of each session.

In the first session, Dr. Jim Murray (UC Davis, USA) reviewed how genetically engineered livestock have been developed for agriculture since the first TARC meeting in 1997. This was closely followed by a talk from Maeve Ballantyne (Roslin Institute, Scotland) about their efforts to engineer resilience to African swine fever into pigs. This disease is rapidly spreading from Africa throughout Eastern Europe. Thus, this type of genetic engineering could be critical for maintaining the health of swine herds. The following talk by Jayne Raper (CUNY, USA), was a natural extension in this session, discussing how genes encoding resistance to trypanosomiasis in non-human primates could be moved into sheep and cattle. The expectation is that such genes are critical for maintaining the health of these herds throughout Africa.

The second session was devoted to new technologies for genome engineering. It started with an excellent review from Bruce Whitelaw (Roslin Institute, Scotland). His review showed how the initial slow progress in generating precisely mutated animals has become much more rapid with the introduction of genome editing. The promise of this technology was soon demonstrated by Mark Tizard (CSIRO, Australia), who described efforts to edit the genome of poultry, and by Bhanu Teluga (University of Maryland, USA), who described his highly efficient CRISPR/Cas targeted genome editing in pigs.

After an afternoon break for hiking, shopping, boating and general fun in Lake Tahoe, there was a late afternoon poster session with submissions from throughout the world. After dinner, the evening session began with a talk from Pablo Ross (UC Davis). Pablo reviewed how pluripotent stem cells have been used to generate targeted livestock, and tantalized the audience with a promise of an upcoming publication describing a new media for growth of pluripotent stem cells from large animals, hopefully capable of generating chimeras and germline transmission. This was closely followed by talks from Franklin West (Univ. of Georgia, USA) and Jorge Piedrahita (NCSU, USA) about the use of stem cells in both pigs and chickens.

The second full day of the meeting was begun with a review by Chris Rogers (Exemplar Genetics, USA) on how genetically engineered livestock have been developed for biomedical models. Simon Bawden (SARI, AU) reported how Huntington’s disease has been recreated in sheep. This was followed by a talk from Lydia Garas (UC Davis, USA) about lysozyme transgenic goats whose milk can be used to prevent and treat intestinal diseases. After a short break, Mingjun Liu (China) described how the sheep FGF5 and MSTN genes have been altered using CRISPR/Cas9 gene editing. The final talk of the morning was from Margareth Capurro (Univ. of Sao Paulo, Brazil), where she captivated the audience with her description of the methods used to gain acceptance for release of GE mosquitoes to reduce the incidence of dengue fever in one Brazilian village. Margareth finished her talk with a most memorable jingle used as a public service announcement!

The Tuesday afternoon session was composed of talks from Eddie Sullivan (SABBiotherapeutics, USA) about the generation of humanized antibodies produced in cows, and from Lissa Herron (Roslin Institute, Scotland) about the isolation of pharmaceutical proteins from avian egg whites. These talks were then followed by an enthusiastic review from Tim Doran (CSIRO, AU) where he surveyed the advances made in engineering of the avian genome. A number of conference attendees added to their notoriety by being listed in his “Hall of Fame”! The final talk on Tuesday, given by Marie-Cecile van de Lavoir (Crystal Biosciences, USA), described the generation of transgenic chickens carrying Cre-recombinase, which can be used to delete selectable markers in vivo.

The final day of the regular conference began with a review by Kevin Wells (Univ. of Missouri, USA) of the regulations governing genetic engineered animals and the food supply. He emphasized that, in the US, while there are regulations that apply, there have not been laws passed that oversee this area, and he called for the preparation of a “white paper” by the experts in the field to advise the US government. His talk was followed by a presentation of the “Glo-fish”@ experience with obtaining US approval given by Alan Blake (Yorktown Technologies, USA). William Muir (Purdue Univ., USA) then presented his statistical model (Hazard Assessment at Critical Control Points, or HAACP) that can assess environmental risk of GE animals based on net fitness of the organism, demonstrating its effectiveness in an experiment on a model organism. He then showed its application to the Aquabounty@ salmon currently awaiting approval, showing that the fear of an accidental release is irrelevant, as the GE salmon would quickly be eliminated from the population.

The next session had talks from Jun Wu (Salk Institute, USA) on the development of pluripotent stem cells, and their use in the pig to generate humanized organs for transplant; and from Hiro Nakauchi (Stanford Univ., USA) on exploiting an “organ niche” by injecting pluripotent stem cells from one organism (rat) into another, deficient organism (Pdx1-/- mouse) to generate a xenogenic pancreas. He is now testing this process in pigs as well.

Attendees were then given another welcome afternoon off to play in the surrounding area, where there is ample opportunity for boating, biking and hiking. This being the final day of the regular conference, everyone truly welcomed this last chance to enjoy the lake and surrounding mountains.

The final session of the meeting began (after another poster session and dinner) with a review given by Heiner Niemann (Hannover, Germany), where he spoke about the use of pigs as xeno-donors for human organs. He described three major hurdles to this scenario, including immune responses, physiological incompatibilities, and the risk of transmitting zoonotic organisms. His own work is an attempt to modify the immune response by humanizing several candidate genes.

The last talk of the meeting was from Alison Van Eenennaam (UC-Davis, USA) about how the technology has progressed but the acceptance of transgenic food animals has not over the past twenty years that TARC meetings have been held. She made an eloquent request that scientists take the time to explain and assure the public that genetic engineering technology can be safe and assist the world with developing a healthy, sustainable food supply. The scientific portion of the meeting then ended with the presentation of the poster award (sponsored by the Roslin Institute) to Dorothea Aumann (Munich, Germany) for her poster on “Analyzing gamma/delta T-cell function in chicken by reverse genetics”. The award presentation was followed by a discussion of how to advance the regulatory environment.

An optional Livestock Industry Day was held the following day, 14 August, 2015, where various company representatives could share their work, interact with attending scientists, and have another enjoyable day in Lake Tahoe. All in all, it was a very informative, interesting, and pleasurable meeting. Granlibakken Conference Center [http://www.granlibakken.com], The UC Davis Department of Animal Science [http://animalscience.ucdavis.edu], Drs. Jim Murray, Elizabeth Maga, Alison Van Eenennaam and Pablo Ross should be commended for their hard work in producing such a successful gathering. The next meeting will be held August 13-17, 2017—please plan on attending!

 

 

Respectfully submitted by:
Jan Parker-Thornburg, with editing from Walter Tsark and Jim Murray

Advertising the TT2014 meeting from your institutions: put one of these Posters!

12th Transgenic Technology (TT2014) meeting, Edinburgh, Scotland, UK, 6-8 October 2014
12th Transgenic Technology (TT2014) meeting, Edinburgh, Scotland, UK, 6-8 October 2014

The next ISTT meeting will be held in Europe this year. The 12th Transgenic Technology (TT2014) meeting, will take place in Edinburgh, Scotland, UK, on 6-8 October 2014, organized by ISTT members Douglas Strathdee (chair), Peter Hohenstein and Bruce Whitelaw, and hosted by three Scottish research institutes and the University of Edinburgh: the Roslin Institute; the Institute of Genetics and Molecular Medicine and the Beatson Institute for Cancer Research. The TT2014 meeting will be followed by the 2-day hands-on workshop “An Introduction to Zebrafish Transgenesis“, on 8-10 October 2014.

An outstanding group of invited speakers have confirmed their participation at the TT2014 meeting. Abstract submissions and application for the ISTT registration awards (for ISTT members) deadlines merge on 30 June 2014. Early bird registration deadline at reduced fees is 31 July 2014. A number of submitted abstracts will be selected for oral presentation on topics including:

  • new technologies in transgenesis
  • pluripotential stem cells
  • targeted nucleases and genome editing
  • models of human disease
  • animal ethics and welfare
  • large-scale phenotyping initiatives
  • animal biotechnology
  • in vivo imaging
  • zebrafish models and transgenesis

Douglas Strathdee and his colleagues have prepared the following collection of eight Posters to advertise the TT2014 meeting, illustrated with beautiful Edinburgh pictures. Please, help us announcing and disseminating the TT2014 meeting by putting one or several of these Posters at your centres, institutions, facilities, departments, universities. The TT meeting is a unique forum occurring every 18 months where to discuss the latest technical developments and applications on animal transgenesis. This is a conference that can’t be missed by anyone interested in this subject! Thanks for helping us advertise the TT2014 meeting!

TT2014 Poster version 1
TT2014 Poster version 1

TT2014 Poster version 1 (A4 format)
TT2014 Poster version 1 (A3 format)

TT2014 Poster version 2
TT2014 Poster version 2

TT2014 Poster version 2 (A4 format)
TT2014 Poster version 2 (A3 format)

TT2014 Poster version 3
TT2014 Poster version 3

TT2014 Poster version 3 (A4 format)
TT2014 Poster version 3 (A3 format)

TT2014 Poster version 4
TT2014 Poster version 4

TT2014 Poster version 4 (A4 format)
TT2014 Poster version 4 (A3 format)

TT2014 Poster version 5
TT2014 Poster version 5

TT2014 Poster version 5 (A4 format)
TT2014 Poster version 5 (A3 format)

TT2014 Poster version 6
TT2014 Poster version 6

TT2014 Poster version 6 (A4 format)
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TT2014 Poster version 7
TT2014 Poster version 7

TT2014 Poster version 7 (A4 format)
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TT2014 Poster version 8
TT2014 Poster version 8

TT2014 Poster version 8 (A4 format)
TT2014 Poster version 8 (A3 format)

Happy New Year and 2013 ISTT Calendar. Renewal campaign launched!

Happy New Year and 2013 ISTT Calendar
Happy New Year and 2013 ISTT Calendar

The International Society for Transgenic Technologies (ISTT) wishes a happy new year and all the best for 2013 to all its members and the entire community of technicians, students and scientists directly or indirectly related to animal transgenesis and animal biotechnology. We have just launched the joining/renewal campaign for 2013 ISTT memberships. We hope that all our 2012 ISTT members will remain with us one more year and that new colleagues will join our Society during 2013. Also, as it has become a new year’s tradition from the ISTT, we have prepared the corresponding 2013 ISTT calendar, this time illustrated with beautiful chimera pictures made by the ISTT Member Benoît Kanzler (Freiburg, Germany), a true artist picturing mice. This calendar, which also contains a number of ISTT and related events indicated, is made freely available to everyone for downloading from the ISTT web site. Looking forward to receive your renewals and requests for registration at the ISTT!

ISTT members chosen for a Panel on Transgenics for the 63rd annual AALAS meeting

63rd AALAS National Meering, Minneapolis, MN, November 3-8, 2012
63rd AALAS National Meering, Minneapolis, MN, November 3-8, 2012

AALAS, or the American Association for Laboratory Animal Sciences is hosting their 63rd Annual Meeting this fall.

For ISTT members planning on attending the AALAS national meeting in Minneapolis MN (USA) in November 3-8, 2012, we would love for you to attend our panel discussion on communications between veterinarians and transgenic facility managers. ISTT members, Jan Parker-Thornberg (MD Anderson) and Aimee Stablewski (Roswell Park Cancer Institute) will be part of the panel discussion along with Rob Taft from the Jackson Laboratory and our veterinarians (Kate Naff, Linda Waterman and Sandra Buitrago, respectively).

AALAS has thousands of applications for presentations so we are fortunate that ours was accepted. This is an ISTT-supported activity. Please come and participate with us.

Melissa Larson (University of Kansas Medical Center), the ISTT member representative before AALAS, will be also taking charge of our ISTT booth this year. Thank you to Melissa, for this great help. If any ISTT member will be attending the meeting, and would like to help Melissa, please email us to volunteer at: aalas@transtechsociety.org

Information about the National AALAS meeting:

Each fall since 1950, the American Association for Laboratory Animal Science (AALAS) has held its annual National Meeting. During the five days of the meeting, members and nonmembers come together to enjoy the workshops, lectures, poster sessions, and exhibits. The program is designed to have topics relevant to the entire membership. Exhibitors have an opportunity to interact with AALAS members from the academic community, research institutions, government organizations, and commercial companies.

The AALAS National Meeting is the largest gathering in the world of professionals concerned with the production, care, and use of laboratory animals. Please see the AALAS meeting website for details on how to register

The International Society for Transgenic Technologies (ISTT) is an AALAS affiliate organization.

We hope to see you there!!