Remembering Laura Pozzi

Laura Pozzi, pioneer of mouse transgenesis in Italy
Laura Pozzi, pioneer of mouse transgenesis in Italy

On August 7th 2016, Laura Pozzi, a pioneer of transgenesis in Italy, passed away at the age of 80.

She was associate professor at the University “La Sapienza” in Rome and during the 80’s of the last century was among the first researchers in Italy to set up, with minimal equipments, often handmade, and with great commitment and personal sacrifice, a laboratory for the generation of transgenic mice. This facility was for a long time one of the few reference points for anyone who wanted to get a transgenic mouse in Italy, so that colleagues jokingly called her “the mother of all the Italian transgenic mice”. Over the years she trained a lot of people on transgenic technologies. On them she had a profound influence and to them she left an irreplaceable legacy of theoretical and practical knowledge that has had a strong impact on their future professional life. Many people who are now operating in the field of transgenesis in Italy, were formed at her school. She was a strict and demanding teacher, but it was so clear to us, her trainees , that her ultimate goal was to provide the best training and formation possible, that we all loved and respected her as a mother and master.

Open-minded, cultivated and great traveler, conversing with her was always pleasant and interesting. Even in private life her main purpose was to be helpful to others: she spent herself as a teacher of Italian language to foreign refugees, like a grandmother she took care of children of friends and neighbours. And all this without bragging and always with her smile and her subtle English humor that she absorbed during a long stay in Great Britain.

After retirement she always remained in touch with her former students and she followed with pleasure and interest, even though she was not a member, the ISTT site using the password of one of us. When, with advancing age, she realized that it was impossible for her to be independent as she wanted and not wanting to be a burden to anyone, with great clarity, serenity and courage she decided that her moment had arrived and left this world, recommending to friends to remember her with a smile.

Elisabetta Mattei, Laura Tatangelo and Isabella Manni

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It is with gread sadness that I learnt from Elisabetta Mattei about the recent passing of Laura Pozzi. She was my first mentor in mouse transgenesis. I was privileged to attend, as the only non-Italian trainee in a small group of five students, a two-weeks intensive practical course on the generation of transgenic mice organized in Siena in 1990, at the headquarters of the pharmaceutical company Sclavo. At the time I was still finishing my PhD in Plant Molecular Genetics, using maize as my experimental model, and I was already preparing myself to begin a future postdoctoral stay in Heidelberg as a mouse geneticist. I landed in Tuscany knowing nothing about transgenic mice and, thanks to the wisdom and teaching abilities of Laura and her team of collaborators, I left Italy with the required solid and robust starting knowledge to work with mice that has travelled with me since then. I went from maize to mice and Laura played a fundamental role in my transitioning between these two experimental models. Throughout the years I remained in touch with Laura and we often commented technical breakthroughs in the field. She was also happy to see the developments and success of the ISTT. I always had her as my first reference in mouse transgenesis. She deserves to be remembered as a most influential person for the mouse transgenics community in Italy and Europe.

Lluis Montoliu

Dolly @ 20

Although she didn’t hit the limelight until February 1997 with a publication in Nature, Dolly-the-most-famous-sheep-in-the-world was born 20 years ago, on 5 July 1996 in The Roslin Institute in Edinburgh. Being the first mammal cloned from adult cells by a team led by Ian Wilmut, Dolly changed the way we look at the up-to-then-supposed irreversibility of development and paved the way for many other forms of reprogramming. Despite suffering from several illnesses early in life, including the well-known arthritis, Dolly was eventually humanely put down on 14 February 2003 when she was found to suffer from a viral form of lung cancer. Dolly’s remains are still on view in the National Museum of Scotland in Edinburgh. Concerns about a potential effect of cloning on premature or accelerated ageing have recently been addressed in a study from the University of Nottingham which shows that 13 cloned sheep, four of which clones from Dolly herself, aged normally.
To celebrate Dolly’s 20th birthday The Roslin Institute is organizing a series of scientific and public events to examine and celebrate her legacy. Public events include a debate in the ‘Cabaret of Dangerous Ideas’ as part of the Edinburgh Festival Fringe, a public lecture and discussion featuring Professor Sir Ian Wilmut, Professor Angelika Schnieke and Noble Prize laureate Professor Shinya Yamanaka in the Surgeon’s Hall Museum in Edinburgh, and a public lecture by Sir Ian at the Virginia-Maryland College of Veterinary Medicine, Blacksburg, Virginia, USA. A number of Dolly-related exhibits will also feature at The Roslin Institute Open Day on the 8 October. There will also be a scientific symposium on 2 September in The Roslin Institute Auditorium that will cover the latest advances in research fields that are linked to Dolly: animal biotechnology, stem cell pluripotency and regenerative medicine. Although the main auditorium is now full, it is still possible to register for a seat in adjacent rooms where the talks will be live-streamed.

Submitted by Peter Hohenstein

Dolly as a lamb with her Scottish Blackface surrogate mother.
Dolly as a lamb with her Scottish Blackface surrogate mother.
Dolly with Professor Sir Ian Wilmut, who led the research which produced her.
Dolly with Professor Sir Ian Wilmut, who led the research which produced her.

 

ISTT at FELASA

13th FELASA Congress, 13-16 June, 2016 – Brussels, Belgium
Respectfully submitted by Boris Jerchow:

Together with Board members Benoît Kanzler and Branko Zevnik, I had the chance to take part in the 2016 edition of the triennial conference of the Federation of European Laboratory Science Associations (FELASA), which took place in Brussels, Belgium, between June 13 and 16. With the kind support of our members Sandra Buhl and Kristin Evans we represented ISTT with a booth. The meeting mainly covered topics important for those involved in laboratory animal science. The conference was divided into six streams:
• Governance, including reports on the transposition of 2010 EU Directive aimed at harmonizing animal welfare standards throughout Europe that has a lot of impact on our work but is still in a phase in which the new regulations are being implemented in daily routines. The stream also addressed active information of the public and ways to conduct an ethical review process. Aurora Brønstad, who also presented at TT2016 in Prague, spoke on the important topic of harm-benefit analysis of animal experiments [1, 2].
• Joint programs across Europe, including education and training, competence management, and 3Rs programs, to name just a few.
• Safety issues with a strong focus on health monitoring of aquatic, amphibian, and rodent species. Moreover, this stream included topics such as occupational health and safety and best working practices but also best practices for husbandry and care, quality of feed, water, and enrichment.

• Common diseases of humans and animals. This stream addressed disease and disease models for metabolic disease, cancer, and also infectious disease including zoonoses. It also included presentations on BLS3 facilities and research with non-human primates.
• Animal well-being emphasized the importance of using the broad understanding of what is going on inside an animal to assess and improve its well-being. The stream contained presentations and discussions about behavioral and neural science, as well as severity assessment, prospectively and also during the time when while animals are being used experimentally by employing clinical signs to recognize pain, suffering distress or lasting harm. I found the latter notably important for our community as we are the ones generating and using genetically altered (GA) animals that may well present with a condition affecting their well-being that should be taken into account before starting the experiment and closely followed during their lifetime. GA animals should also be monitored for unforeseen effects that may compromise their well-being and measures to alleviate pain, suffering, distress or lasting harm should be taken

whenever possible.
• Animal models and animal experiments. For this session I had been asked to convene a session with the title “Genetic modification – technologies and pitfalls”. Together with Ann van Soom, Tom Vanden Berghe and Lluis Montoliu, we informed the audience about the latest technologies in the field, the relevance of the non-coding genome, the threat of passenger mutations, and epigenetic effects. Other sessions in this stream discussed experimental design and reporting, how to process, share, and review acquired as well as existing data, imaging techniques, and various experimental paradigms.
In general there was a strong focus on animal welfare, especially on refinement of procedures, which was present over all streams and sessions. Much of the conference was under the influence of the still ongoing implementation of the regulations of the EU Directive on animal experimentation. Due to the importance of European scientific activities in the world, the pressure on scientific publishers to adopt higher animal welfare and reporting standards, and tight cooperation with countries outside the EU, namely the United States, these regulations will impact on animal experimentation worldwide. The same is true for health standards where FELASA guidelines have already become a gold standard. Although FELASA Conferences draw an audience quite distinct from our TT Meetings, there is a small but important overlap in interest. I am convinced that the ISTT should keep on striving to foster a vivid exchange with FELASA and the laboratory animal science community.

1. Bronstad, A., et al., Current concepts of Harm-Benefit Analysis of Animal Experiments – Report from the AALAS-FELASA Working Group on Harm-Benefit Analysis – Part 1. Lab Anim, 2016. 50(1 Suppl): p. 1-20.
2. Laber, K., et al., Recommendations for Addressing Harm-Benefit Analysis and Implementation in Ethical Evaluation – Report from the AALAS-FELASA Working Group on Harm-Benefit Analysis – Part 2. Lab Anim, 2016. 50(1 Suppl): p. 21-42.

ISTT Booth at FELASA
Benoit Kanzler, Sandra Buhl and Boris Jerchow
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Pens and futbol at the ISTT Booth

ISTT at the 55th annual CALAS meeting

On June 11-14, 2016, the ISTT hosted a booth at the 55th CALAS annual symposium that was held in Toronto, Ontario, Canada. CALAS, the Canadian Association for Laboratory Animal Science, is a national association dedicated to providing high quality training and educational resources to animal care professionals across Canada. CALAS has almost 1,000 members and supports a diverse group of animal care attendants, animal health technicians, and veterinarians. It provides training and certification programs recommended by the Canadian Council on Animal Care (CCAC). The theme of this year symposium attended by 400 participants was: “The dirt on germs: the good, the bad, the unknown”.

Marina Gertsenstein attended the meeting, both to represent the ISTT at the booth and to organize a workshop on Current Technologies in Mouse Genome Manipulations at The Centre for Phenogenomics (TCP). At the booth, ISTT information flyers describing the benefits of the membership and pens with ISTT logo were handed out. Several attendees expressed interest in becoming the members of ISTT.

The highlight of the scientific session was the talk of Dr. Kevin C. Kain, Canada Research Chair in Molecular Parasitology. In his keynote address he described his research on host-parasite interactions responsible for major global health threats such as malaria and HIV and first-hand experience with the clinical problems. This leading researcher is developing effective therapeutic interventions using animal models to determine the molecular basis for clinical outcomes of life-threatening infections and to translate this knowledge into novel therapeutic interventions.

Meeting Report respectfully submitted by Marina Gertsenstein

CALAS logoISTT @CALAS2016CALAS meeting info

Will the novel CRISPR/Cas9 technology for the generation of genetically modified animals increase the number of animals used and lead to a shift in the species used? Statement of the ISTT’s 3Rs Committee

The CRISPR/Cas9 technology emerged only recently. Still it is already obvious, that it makes the generation of genetically modified organisms more efficient than with conventional techniques. Moreover, species that were recalcitrant to those manipulations in the past are now amenable to genome editing.

What we initially saw in the rodent, especially the mouse research community, was a rush into the technique, where many scientists wanted to employ the new technology (me too). While there were a number of reports on off-target effects, it is now clear that off-target genome alterations are rarely found in an in vivo situation (Seruggia et al., 2015; Shen et al., 2014). Moreover modified nucleases are meanwhile available with no detectable off-target effects (Kleinstiver et al. 2016).

In the most widely used model organism in biomedical research, the mouse, the number of animals needed to generate a genetically modified line remains approximately the same with CRISPR/Cas9 as compared to conventional techniques. However, the ease at which new mutations are generated with the new system might lead to increased numbers of novel lines being produced (Williams et al., 2016). Moreover, numbers are also increasing for other species, especially zebrafish and rats (Auer and Del Bene, 2014; Hwang et al., 2013; Wang et al., 2015). As with all novel genetically modified lines, after their generation CRISPR/Cas9 generated animals need to be bred for experiments and are kept on the shelf for considerable time. Thereby, the increase in the number of lines generated will translate into larger numbers of additional animals bred.

At this time, conventional techniques still have their justification in the lab since precise modification of DNA via homologous recombination, especially with large constructs, is not as robust as necessary via CRISPR/Cas9. However, one has to take into account that this is a very recent technology, whilst work to improve the generation of genetically modified mice via homologous recombination in embryonic stem cells has been ongoing for more than 25 years. We therefore may see extensive improvements over the coming years once we gain a better understanding of the underlying mechanisms. With improvements in targeting efficiencies there could be opportunities for further reductions in the number of mice used for the generation of the animal model to be studied: In many cases mutations are still introduced into ES cells that are used for the generation of live mice. Even though culture conditions have been improved over the years, cell clones show a fairly high degree of aneuploidy so that multiple clones have to be injected to generate chimeric offspring that transmit the mutation to their progeny. Due to the non-chimeric nature of CRISPR/Cas9, mutated individuals will in most cases transmit. Therefore an improved ratio of mutant to wildtype offspring could directly reduce the number of animals produced. However, the degree of mosaicism could counteract this reduction. CRISPR/Cas9 technology has the potential that several mutations can be introduced on different alleles at the same time, even homozygously, once the mosaicism issue is solved. Alternatively, complex mutations can be generated by adding additional mutations in pronuclear stage embryos of mutant backgrounds. With a sufficient increase in efficiency, this promise does appear realistic (Williams et al., 2016). This would mean that instead of producing a plethora of unwanted mice with unwanted genotypes in the process of breeding compound mutants, one could proceed right to the desired combination of mutated alleles.

We expect to see an increase in the number of species that will undergo complex targeted genetic manipulation. Still, a significant increase in laboratory animal numbers is not expected. The research infrastructure has been developed and is still being developed for large scale mouse breeding. The breeding of rats takes up much more space and only a few centers have the infrastructure for larger animals. The applicability of the technology to non- human primates means they are also more often nowadays used for transgenic animal research, especially in countries where such research is not strictly regulated. This is another issue of public concern and will require an ethical evaluation process beyond the scope of the 3R approach. CRISPR/Cas9 technology is widely used to genetically manipulate zebrafish. An increase in zebrafish numbers similar to what is foreseen in the mouse can be expected. On the other hand there will be projects where the zebrafish is now amenable to the introduction of complex targeted genetic manipulation and can therefore replace the mouse as a model – a clear refinement in accordance with the 3Rs.

In summary, the ISTT’s 3Rs Committee acknowledges that the advent of the CRISPR/Cas9 technology has the potential to significantly increase the number of animals used and range of species genetically modified. However the committee believes that existing limits on space and other associated resources will inhibit the realization of this at the current time. In the long run, after the technology has been developed further and improved, the Committee is hopeful that opportunities for reducing the number of animals that are bred but not used will be realized.

Boris Jerchow, Chair
On behalf of the ISTT’s 3Rs Committee Berlin in March 2016

Auer, T.O., and Del Bene, F. (2014). CRISPR/Cas9 and TALEN-mediated knock-in approaches in zebrafish. Methods 69, 142-150.
Hwang, W.Y., Fu, Y., Reyon, D., Maeder, M.L., Tsai, S.Q., Sander, J.D., Peterson, R.T., Yeh, J.R., and Joung, J.K. (2013). Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol 31, 227-229.

Kleinstiver, B.P., Pattanayak, V., Prew, M.S., Tsai, S.Q., Nguyen, N.T., Zheng, Z., and Joung, J.K. (2016). High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off- target effects. Nature, 529, 490-495.
Seruggia, D., Fernandez, A., Cantero, M., Pelczar, P., and Montoliu, L. (2015). Functional validation of mouse tyrosinase non-coding regulatory DNA elements by CRISPR-Cas9- mediated mutagenesis. Nucleic acids research 43, 4855-4867.

Shen, B., Zhang, W., Zhang, J., Zhou, J., Wang, J., Chen, L., Wang, L., Hodgkins, A., Iyer, V., Huang, X., et al. (2014). Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects. Nature methods 11, 399-402.

Wang, L., Shao, Y., Guan, Y., Li, L., Wu, L., Chen, F., Liu, M., Chen, H., Ma, Y., Ma, X., et al. (2015). Large genomic fragment deletion and functional gene cassette knock-in via Cas9 protein mediated genome editing in one-cell rodent embryos. Scientific reports 5, 17517. Williams, A., Henao-Mejia, J., and Flavell, R.A. (2016). Editing the Mouse Genome Using the CRISPR-Cas9 System. Cold Spring Harbor protocols 2016, pdb top087536

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

LASA Winter Meeting – Genome Editing Session

Meeting Report by Mary-Ann Haskings, 25 November, 2015

It was a cold, sunny day in Brighton, UK for the LASA Winter meeting and we were pleased to see several ISTT members attending the meeting. The talks reflected on recent work involving genome editing across a breadth of species: mouse, zebrafish, opossum and discussion of use in humans.

There were some common themes across the talks and some newer approaches such as testis electroporation highlighted. The occurrence of mosaicism was discussed lengthily. The use of the NHEJ inhibitor SCR7 was another hot topic, with multiple speakers reporting that their experience suggested that there was little gain in using it. Comparisons were shown between Cas9 mRNA and protein; also the use of transgenic mice overexpressing Cas9 was reported. The possibility of reproducing better models of multi locus disease was recognised by several presenters.

The afternoon session focused on the ethical considerations of the technology, with the reminder that while it may well cause a reduction in numbers as we refine the production of genetically altered animals, the ease and efficiency of the technology may actually lead to a rise in the numbers of animals being used.

We closed with a round table discussion giving the audience the opportunity to ask any questions of our speakers. We need to thank all our speakers and our fantastic chairs Brendan Doe and Sarah Hart Johnson who helped the day to run so smoothly.

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

Tenth Transgenic Animal Research Conference, Tahoe City, California (USA), 9 – 13 August, 2015

TARC_2015blog

Plan to attend the 10th Transgenic Animal Research Conference (TARC X) in August of 2015. At this international meeting you will learn the latest developments in the field of non-murine transgenic animals. In celebration of the 10th conference in this series the program will contain nine review talks, to be published in a special issue of Transgenic Research. Once again the conference will be held at the beautiful Granlibakken Resort and Conference Center, high in the Sierra Nevada Mountains adjacent to beautiful Lake Tahoe. This meeting is co-sponsored by the ISTT.

Rooms are limited, so plan to register early. The conference web site opened February 1, 2015 for registrations and submission of poster abstracts. The following list of speakers confirms again that this is conference not to be missed. Additionally, in conjunction with Recombinetics, Inc there will be a special one day program on August 13th for the livestock, poultry and aquaculture industries on the application of GE animals. A list of confirmed speakers and topics, as well as additional information, registration and poster submission forms may be found on the conference web site (http://conferences.ucdavis.edu/transgenic). We invite you to join us for this interesting and important conference and learn more about the genetic future of the livestock industry.

Confirmed Speakers:

Reviews

  • Elizabeth Maga/Jim Murray (UC Davis) GE livestock for agriculture
  • Chris Rogers (Exemplar Genetics) GE livestock for biomedical models
  • Heiner Niemann (Hannover) Xenotransplantation
  • Tim Doran (CSIRO, Australia) GE Poultry
  • Pablo Ross (UC Davis) iPS/Stem cells
  • Jun Wu (Salk Institute) Organ complementation
  • Bruce Whitelaw (Edinburgh) Gene editing/gene targeting
  • Luciana Bertolini (Brazil) Production of pharmaceuticals
  • Kevin Wells (Missouri) Regulation of transgenic animals

Additional speakers and topics

  • Maeve Ballantyne (Roslin) African swine fever resistant pigs
  • Simon Bawden (Australia) Huntington’s disease sheep model
  • Jayne Raper (New York) Cattle resistant to trypanosomiasis
  • Lydia Garas (UC Davis) Effects of lysozyme milk on intestinal health
  • Jorge Piedrahita (NC State) SCID pigs
  • Margarthe Cupurra (Sao Paulo) GE mosquitos to control dengue fever
  • Lissa Herron (Roslin) Pharmaceuticals from eggs
  • Eddie Sullivan (SABBiotherapeutics) Targeting emerging infectious diseases through animal biotechnology
  • Hiro Nakauchi (Stanford) Interspecies blastocyst complementation

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 tgcourse2015@pasteur.edu.uy
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 tgcourse2015@pasteur.edu.uy
SYMPOSIUM fee is U$S 100.

SAVE THE DATES!
Deadline for COURSE applications is June 28th
Deadline for SYMPOSIUM registrations is July 19th
For any further information contact: tgcourse2015@pasteur.edu.uy
www.pasteur.edu.uy/index.php/en/futurecourses