Archive for the ‘knockout’ Category

Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)

Tuesday, July 22nd, 2014
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Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)

Manipulating the Mouse Embryo. A Laboratory Manual (4th edition, 2014)

A bit more than 10 years later, from the last edition (3rd, 2013) published, the classical manual, the “Bible” in our field has been renewed, updated, upgraded and recently published in its fourth edition. The new “Manipulating the Mouse Embryo. A Laboratory Manual. 4th edition, 2014” has just been released, published by Cold Spring Harbor Laboratory Press and edited by Richard Behringer, Marina Gertsenstein, Kristina Vintersten Nagy and Andras Nagy. The editors, the same team that produced the 3rd edition of this useful manual, should be praised once again for a brilliant work done, accommodating the latest techniques in mouse embryo manipulation, while not forgetting the traditional procedures that must be learnt properly by anyone joining this field of animal transgenesis for the first time.  The editors counted with the help of many additional collaborators, who provided materials and protocols, including many ISTT members, as Marina and Andras are, such as:  Wojtek Auerbach, Ralph Brinster, Jorge Cabezas, Tracy Caroll, Lluis Montoliu, Naomi Nakagata, Jan Parker-Thornburg, Shirley Pease and Thomas Saunders, just to name a few of the many colleagues that shared their expertise and contributed to this 4th edition of the CSHLP manual.

This fourth edition is dedicated to Anne McLaren (1927-2007), one of the brave and gifted pioneer in mouse genetics and embriology, and whom do we all owe a number of standard procedures that we routinely apply in a transgenic laboratory, such as mouse embryo culturing, pseudopregnancy and embryo transfer.

The structure of this 4th edition manual follows that of the 3rd edition, in the sense that chapters and methods are grouped logically and functionally, from a very good summary on mouse genetics and mouse embryo development, to mouse colony management, and followed by a variety of main sections dealing with in vitro and in vivo work, surgical procedures, the production of transgenic mice by pronuclear microinjection, derivation of ES cell lines, using ES cells to generate germline-transmitting chimeras, genotyping protocols, parthenogenesis and nuclear transfer, assisted reproduction techniques, cryopreservation methods, techniques for visualizing gene products and finishing by a chapter devoted to setting up a micromanipulation laboratory and the usual and convenient appendix with receipts for the common buffers and solutions.

Besides this excellent updated version of the CSHLP Mouse Manual, there are several books published on this subject, including the ISTT manual (Springer, 2011) edited by Shirley Pease and Thomas Saunders, and all are worth reading and having them around for consultation, since each one brings their own contribution, their view and their solutions for the adequate learning of how to best manipulate animal embryos, and not all methods and protocols are covered in a single book, hence they are largely complementary. I will be soon reporting in this ISTT blog about additional books recently published in this field.

 

TT2014 abstracts: submission deadline is approaching (30 June)

Tuesday, June 17th, 2014
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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
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!

Highlights of the TT2014 meeting in Edinburgh: a conference you can’t miss!

Thursday, June 5th, 2014
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The TT2014 meeting in Edinburgh (6-8 October 2014)

The TT2014 meeting in Edinburgh (6-8 October 2014)

This year’s ISTT main activity is the 12th Transgenic Technology conference, the TT2014 meeting, which will be held in Edinburgh, Scotland, UK, on 6-8 October 2014, followed by a 2-day hands-on workshop on basic zebrafish transgenesis techniques. The ISTT promotes the TT meetings every 18 months, these being the most important activity of our Society. This year, the local Organizers and advisory committees, commanded by Douglas Strathdee, need to be praised for preparing a most appealing and interesting program, addressing hot topics, current and most up-to-date issues actively discussed nowadays by the transgenic animal community. Talks that will be presented by the most active and prestigious scientists in our field.

Why you shouldn’t miss the TT2014 meeting?

  • If you are interested in the new transgenic methods associated to nucleases (ZFNs, TALENs and CRISPRs-Cas9) there will be plenty of  interesting talks where these new fantastic tools will be presented and discussed, directly by the key players in this rapidly-evolving field, including: Rudolf Jaenisch, William Skarnes, Angelika Schnieke, Kai Schönig, Ignacion Anegon, Pawel Pelczar, Francis Stewart, Keith Joung and Feng Zhang. And, most likely, these techniques will be referred and cited in many additional talks too, including the round table discussion about the future of transgenic core facilities, chaired by James Bussell.
  • If you are interested in ES cell biology and in innovative uses of ES cells and associated technologies there will be unique talks delivered by Jos Jonkers, Austin Smith, Ian Chambers, Janet Rossant and Alex Joyner
  • If you are interested in phenotyping your mouse animal models there will be fantastic talks delivered by  Jacqueline White, Stephen Murray, David Adams, Daniel MurphyAnna-Katerina Hadjatonakis and Vasilis Ntziachristos
  • If you are interested in non-rodent, large mammals and birds, animal models there will be great talks by James Murray, Angelika Schnieke, Mike McGrew and Adrian Shermann
  • If you are interested in rats there will be compelling talks by Kai Schönig and Ignacio Anegon
  • If you are interested in zebrafish animal models there will be fascinating talks by Stephen Ekker, Koichi KawakamiKeith Joung and Elizabeth Patton
  • If you are interested in animal welfare and 3Rs, in the best use of our laboratory animals, there will be captivating talks by Peter Hohenstein, Sara Wells and Jan-Bas Prins.

Therefore, there will be really engaging talks interesting to everyone in our field. This is why you shouldn’t miss this great and unique opportunity!.

Register now for the TT2014 meeting. Submission of abstracts will be accepted up to June 30. Early-Bird registration at reduced fees will be promoted up to July 31. ISTT members are entitled to reduced fee registration.

See you all in Edinburgh in October!

 

Meeting report: Promoting the international exchange of mouse mutant resources

Monday, May 12th, 2014
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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. 2012Mishra 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

Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June

Sunday, May 11th, 2014
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Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June

Submit your abstract(s) to the TT2014 meeting in Edinburgh: 30 June

The submission of abstracts for the 12th Transgenic Technology (TT2014) meeting, to be held in Edinburgh (Scotland, UK), on 6-8 October, is OPEN. 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), and, 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
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. All presented Posters at the TT2014 meeting will be entitled to the Best Poster Awards, generously sponsored by Charles River.

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

A report on the 7th European Short Course on Laboratory Animal Science in Strasbourg, organized by Charles River

Friday, February 14th, 2014
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A report on the 7th European Short Course on Laboratory Animal Science in Strasbourg, organized by Charles River

A report on the 7th European Short Course on Laboratory Animal Science in Strasbourg, organized by Charles River

The 7th European Short Course on Laboratory Animal Science, organized by Charles River, just closed in Strasbourg, France, after three days of interesting presentations and discussions at the intersection between animal welfare, animal experimentation, current guidelines and legislation, biomedical research from academia and industry and society perception on these topics. The Organizers should be praised for the selection and variety of topics, as well as for the smooth and pleasant running of the entire course, which included an enjoyable visit to an old typical cellar from the Alsace region along with a wine-testing Gala dinner.

Several ISTT members participated in this event, including organizers (Cyril Desvignes, Jean Cozzi), members of the steering committee (Johannes Wilbertz), invited speakers (Belén Pintado, Yann Herault, Ignacio Anegon, Lluís Montoliu), and participants (Marcello Raspa, Ferenc Erdelyi, Gabor Szabo,…) among other.

During this course, the recent EU Directive 2010/63 on the protection of animals used in research and its implication on the use of animals in biomedical research and policies throughout Europe was discussed, from different angles, by Magda Chlebus, Gill Fleetwood, Thierry Decelle, Patri Vergara and Belén Pintado. Topics covered included the new training courses and competencies to work with experimentation animals in Europe, the animal-welfare bodies and the current understanding of the 3R’s paradigm. Javier Guillén compared, side by side, the new EU Directive with the current Guide in the US and highlighted their many coincidences, suggesting that a combined use of both documents would be ideal for the adoption of successful animal care and use programs. Jan-Bas Prins, current president of FELASA, presented his view of the field of laboratory animal sciences, before the implementation of this new EU Directive, as an opportunity and a positive challenge to interface and exchange knowledge with many other players involved.

Health monitoring programs, rodent microbiologic surveillance, methods employed to detect all these pathogens robustly in laboratory animal facilities and the updated recommendations from FELASA, recently published in Laboratory Animals, were presented by William Shek, Guy Mulder, Stéphanie Durand and Axel Kornerup Hansen. Operational and technical aspects of animal facilities were discussed by Alberto Gobbi and Peter Dockx, whereas the issues related with occupational health and safety program evaluations were presented by Jann Hau.

Examples of the use of rodent animal models in biomedical research, in academia, by James Di Santo and Andrea Bertotti, as well as in the industry, by Joyce L. Young, were discussed. The importance of genetic quality in mouse research as well as the complexity of mouse genome and the impact of the genetic background on phenotypes was presented by Charles Miller and Lluís Montoliu, respectively. The procedures conducted at the International Mouse Phenotyping Consortium (IMPC) as well as the challenges they encountered during the deployment of this impressively large enterprise were presented and discussed by Sara Wells and, by the local representative, Yann Herault, Director of the French Mouse Clinic, ICS, in Strasbourg, who delivered the closing talk.

The newest technologies in stem cell biology and animal transgenesis were also present at this 7th Short Course. Hongkui Deng summarized the most innovative approach he devised to prepare induced-pluripotent cells from somatic cells, using a cocktail of four chemicals, four molecules that mimicked the induction signals described by Shinya Yamanaka. The new logics for the production of targeted genetic modifications, using editing or engineered nucleases (Meganuclease, ZFNs, TALENs, CRISPRs) in mice and rats was presented by Ralf Kuehn and Ignacio Anegon, respectively.

The choice of rodent anaestesia protocols was discussed by Aurelie Thomas, whereas the various methods for euthanasia in rodents were presented by Huw Golledge. On the last day, Aurora Bronstad summarized the work done at the AALAS-FELASA joint working group on harm-benefit analysis, whereas Katrina Gore highlighted the need for more robust analytical procedures in research protocols involving animal experimentation, in order to optimize the rate of success of pre-clinical drugs.

In summary, the 7th Edition of this biennial Charles River Short Course on Laboratory Animal Science in Europe, attended by some 120 participants, was an excellent opportunity to update information related to animal welfare, EU legislation and transposition difficulties in various countries, newest technologies, mouse genomics and genetics, large mouse consortia and numerous important topics that are relevant for animal facility managers, researchers, veterinarians and anyone else interested in the best use of animals in experiments, according to current laws and recommendations.

More than 27,000 messages on animal transgenesis available to ISTT members through ISTT_list and tg-l archives

Sunday, February 9th, 2014
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More than 27,000 messages on animal transgenesis available through ISTT_list and tg-l archives

More than 27,000 messages on animal transgenesis available through ISTT_list and tg-l archives

One of the most important assets of the International Society for Transgenic Technologies (ISTT), is the amount of information on animal transgenesis accummulated through the archives of the ISTT_list and tg-l email lists. Currently, more than 27,000 messages are fully available to ISTT members, conveniently organized in searchable and dynamic archives. The traditional transgenic-list (tg-l), operative since 1996 and offered from the ISTT web server since the end of 2011, has distributed over 22,000 messages since then, whereas the ISTT_list, associated and born with our Society in 2006, has disseminated some 5,000 messages, discussing both lists on almost each and every topic, issue or situation related directly or indirectly with animal transgenesis. All this endless information resource is fully available to ISTT members, through powerful search engines. Non-ISTT members subscribing to tg-l have access only to the most recent messages distributed through the tg-l, using the simple search engine, which allows simple searches and outputs the 50 most recent messages discussed on the subject of interest. In contrast, ISTT members have access to more sophysticated searching engines and the output always contains all messages archived on the matter investigated.

Obtaining granted access to these rich sources of information is very easy and cheap. Simply apply for ISTT membership! Submit now your application to become a member of the ISTT and you will get immediate and full access to all these messages.

Updated scientific and workshop programmes for the TT2014 meeting in Edinburgh

Friday, January 10th, 2014
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Upades scientific and workshop programmes for TT2014 meeting in Edinburgh: Please, register today!.

Upades scientific and workshop programmes for TT2014 meeting in Edinburgh: Please, register today!.

The scientific and zebrafish transgenesis hands-on workshop programmes prepared for the 12th Transgenic Technology (TT2014) meeting, to be held in Edinburgh (Scotland, UK), on October 6-8 (workshop on October 8-10) 2014,  have been recently updated by the Organizers, chaired by Douglas Strathdee (Glasgow, UK). These rewarding updates further increased the already high quality and interest for this popular conference series, promoted from the International Society for Transgenic Technologies (ISTT), the most important forum where to discuss the state-of-art of animal transgenic technology, to share new developments, to review the deployment of the new methods that have recently being devised and, in summary, an excellent arena where to easily meet, face-to-face, the most relevant key-players in the field while providing a wonderful excuse to gather and ex-change experiences with the entire ISTT family of members.

The updated list of confirmed invited speakers attending the TT2014 meeting (6-8 October 2014) includes:

  • David Adams, Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK
  • Ignacio Anegon, Center for Research in Transplantation and Immunology, Nantes, France
  • James Bussell, Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK
  • Ian Chalmers, MRC Centre for Regenerative Medicine, The University of Edinburgh, UK
  • Stephen Ekker, Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, USA
  • Anna-Katerina Hadjatonakis, Developmental Biology Program, Sloan-Kettering Institute, New York, USA
  • Peter Hohenstein, The Roslin Institute and Royal Dick School of Vetinary Studies & MRC IGMM, University of Edinburgh, UK
  • Rudolf Jaenisch, Whitehead Institute for Biomedical Research, Nine Cambridge Center Cambridge, USA
  • Jos Jonkers, Division of Molecular Pathology, The Netherlands Cancer Institute, Amsterdam, The Netherlands
  • Keith Joung, Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA
  • Alexandra Joyner, Developmental Biology Program, Sloan-Kettering Institute, New York, USA
  • Koichi Kawakami, Division of Molecular and Developmental Biology, National Institute of Genetics, Shizuoka, Japan
  • Michael McGrew, Division of Developmental Biology, The Roslin Institute and Royal Dick School of Veterinary Studies, University of Edinburgh, UK
  • Daniel J Murphy, Beatson Institute for Cancer Research, University of Glasgow, UK
  • James Murray, Department of Animal Science and Department of Population Health and Reproduction, University of California, Davis, California, USA
  • Stephen Murray, The Jackson Laboratory, Bar Harbor, Maine, USA
  • Lluis Montoliu, ISTT President, Organising Committee, National Center of Biotechnology (CNB), CSIC, Madrid, Spain
  • Vasilis Ntziachristos, Technische Universität Mu?nchen, Munich, Germany
  • Elizabeth Patton, MRC Human Genetics Unit & MRC IGMM, University of Edinburgh, UK
  • Pawel Pelczar, Institute of Laboratory Animal Science, Zürich, Switzerland
  • Jan-Bas Prins, Leiden University Medical Centre, The Netherlands
  • Janet Rossant, The Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada (ISTT Prize)
  • Angelika Schnieke, Livestock Biotechnology, WZW Center of Life Science, Freising-Weihenstephan, Germany
  • Kai Schönig, Central Institute of Mental Health, Heidelberg University, Mannheim, Germany
  • William Skarnes, Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK
  • Austin Smith, Wellcome Trust-Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK
  • Francis Stewart, Biotechnology Center of the TU Dresden, Germany
  • Sara Wells, MRC Harwell, Oxfordshire, UK
  • Jacqueline White, Wellcome Trust Sanger Institute, Hinxton, Cambridge UK

The updated list of confirmed invited speakers & instructors attending the hands-on zebrafish transgenesis workshop taking place immediately after the TT2014 meeting (8-10 October 2014) includes:

  • Liz Patton, MRC Institute of Genetics and Molecular Medicine, The University of Edinburgh
  • Carl Tucker, Biomedical Research Resources, The University of Edinburgh
  • Tim Czopka, Centre for Neuroregeneration, The University of Edinburgh
  • Koichi Kawakami, Division of Molecular and Developmental Biology, National Institute of Genetics, Shizuoka, Japan
  • Stephen Ekker, Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, USA
  • Keith Joung, Department of Pathology, Massachusetts General Hospital and Harvard Medical School
  • Henry Roehl, Department of Biomedical Science, The University of Sheffield
  • Robert Kelsh, Centre for Regenerative Medicine and Department of Biology and Biochemistry, The University of Bath
  • Martin Denvir, The University of Edinburgh/British Heart Foundation Centre for Cardiovascular Science, The University of Edinburgh
  • David Lyons, Centre for Neuroregeneration, The University of Edinburgh
  • Dirk Seiger, Centre for Neuroregeneration, The University of Edinburgh
  • Karthikeyani Paranthaman, MRC Institute of Genetics and Molecular Medicine, The University of Edinburgh

Abstracts: All TT2014 participants are encouraged to submit their work as an abstract for poster presentation at the TT2014 meeting. 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. 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. 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

Registration for both the TT2014 meeting and the zebrafish transgenesis workshop are OPEN. Registration for the TT2014 meetings starts at 265 UK Pounds for technician/student ISTT members and progressively increases for the rest of categories of delegates. ISTT members are always entitled to reduced registration fees. Registration for the zebrafish transgenesis workshop is independent, with an extra cost of 275 UK Pounds , and only open to delegates that have also registered to attend the TT2014 meeting. The early bird reduced registration fees are operative until July 31, 2014. Thereafter, registration will be progressively become more expensive. Hence,  please register by July 31, 2014 to benefit from reduced registration fees.

ISTT Registration Awards: Application to ISTT registration awards for the TT2014 meeting is OPEN. A minimum of six registration awards for ISTT members will be sponsored by the International Society for Transgenic Technologies (ISTT). Applications should be sent, along with the registration confirmation and the requested additional documents to istt@transtechsociety.org by June 30, 2014. The ISTT will pay the Registration Fee of all applicants selected for an award. Please note that applicants not selected for an award are required to pay the coresponding registration fee. Please note the Award covers registration fees and attendance at all social events, however, does not cover travel, accommodation expenses or attendance at pre meeting events. Award decisions will be communicated by July 15, 2014 and awardees will receive a diploma at the TT2014 Meeting. Deadline for submitting application for ISTT Registration Awards for TT2014: 30 June 2014. Registration Award decisions will be communicated by 15 July 2014.

Looking forward to meeting you all in Edinburgh!

 

Cell-permeable Cre recombinase for rapid conversion of EUCOMM/KOMP-CSD alleles

Wednesday, November 13th, 2013
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Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase. (Figure 1 from Ryder, Doe et al. Transgenic Research, 2013 Nov 7. DOI: 10.1007/s11248-013-9764-x)

Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase. (Figure 1 from Ryder, Doe et al. Transgenic Research, 2013 Nov 7. DOI: 10.1007/s11248-013-9764-x)

Our colleagues from the Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, UK, most of them ISTT members, have just published a very interesting paper in Transgenic Research, the scientific journal associated with the International Society for Transgenic Technologies (ISTT),  describing the use of a cell-permeable Cre recombinase for the rapid conversion of EUCOMM/KOMP-CSD alleles directly in mouse embryos, hence avoiding the traditional breeding of mice produced with the original tm1a alleles with Cre-transgenic mouse lines to produce non-conditional knockout alleles (tm1b allele). This innovative procedure saves time, money, and, most importantly, many animals, thus contributing to animal welfare.

Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase.
Ryder E, Doe B, Gleeson D, Houghton R, Dalvi P, Grau E, Habib B, Miklejewska E, Newman S, Sethi D, Sinclair C, Vyas S, Wardle-Jones H; Sanger Mouse Genetics Project, Bottomley J, Bussell J, Galli A, Salisbury J, Ramirez-Solis R.
Transgenic Res. 2013 Nov 7

Workshop report: animals bred, but not used in experiments

Wednesday, October 23rd, 2013
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Workshop:  “Animals bred, but not used in experiments”, October 18-20, 2013, Hotel Duin & Kruidberg, Santpoort, the Netherlands (Picture kindly provided by Fernando Benavides)

Workshop: “Animals bred, but not used in experiments”, October 18-20, 2013, Hotel Duin & Kruidberg, Santpoort, the Netherlands (Picture kindly provided by Fernando Benavides)

Workshop: “Animals bred, but not used in experiments”, October 18-20, 2013, Hotel Duin & Kruidberg, Santpoort, the Netherlands.

Experiments in biomedical science use large numbers of laboratory animals. It is a fact that to provide these animals, regularly more animals are bred than are finally used in the experiments planned. The Ministry of Economic Affairs as the competent body of the Netherlands had asked Prof. Coenraad Hendriksen and Dr. Jan-Bas Prins to organize a workshop to identify the reasons for the breeding of surplus animals and to devise recommendations as to how the number of animals that are bred but not used can be reduced to a minimum.

A number of experts from different fields of laboratory animal science were invited for a two day workshop to the Hotel Duin & Kruidberg in Santpoort, a town close to Amsterdam, to discuss these issues and to develop a paper for the Dutch authorities. Obviously, many of the laboratory animals bred are genetically altered (GA) animals. Moreover, techniques to cryopreserve GA animal lines could be a means to reduce the number of animals that are bred. The invitation was therefore extended to the ISTT to send a representative to take part in this workshop.

Here, I will give a short summary of the topics that have been discussed and of the outcomes. However, I refer you to the final report of the workshop, parts of which have been developed within individual small workgroups and will be put together into a final document by the kind efforts of Coenraad and Jan-Bas. I will inform you immediately upon the publication of this report.

A topic central to the discussion was the identification of reasons for the production of animals that are then not used in experiments. A major reason for this is the production of unwanted sexes and unwanted genotypes. The participants agreed that good planning can considerably reduce the number of surplus animals. At the same time, resources can be saved and either used for additional experiments or for cost reduction. However, breeding schemes with multiple alleles, as well as the organization of a facility, can be complex. A strong need for counseling as well as education of users of laboratory animals was identified, to make them competent to plan accordingly. The centralization of the breeding colonies under the responsibility of the facility management was discussed as a possibility to streamline breeding strategies. On the other hand, for the time being, this does not seem to be feasible for very many facilities. Local Animal Welfare Committees should evaluate local SOPs and develop a catalogue of best practices to help keep surplus animals to a minimum. GA animal lines should be cryopreserved immediately after their creation when there is no need to breed extra animals for this purpose and when animals from test rederivations can be used for experiments or for the breeding colony. Thereby, the lines are protected from disaster and from genetic drift at the same time, live mice can be terminated at any time, and the lines can be easily shipped to collaborators. Lines should be made available to collaborators as early as possibly to avoid generating the same line at different places. In case expertise for cryopreservation is lacking, lines can be donated to repositories like EMMA where they are cryopreserved free of charge. Investigators should always consider sharing lines with the scientific community through such repositories.

A second important topic discussed during the workshop was the use of new technologies for the generation of GA animals as well as for their experimental analysis. New lines should be directly generated on the desired background. In case backcrossing is needed, speed congenic strategies should be used to reduce the number of animals needed during that process. Technologies utilizing the targeting of nucleases to the locus of interest (ZFNs, TALENs, CRISPER/Cas9) promise to eventually allow the generation of GA lines with reduced numbers of animals directly on the desired background. Complex strategies for the generation of customized animals for specific experiments were presented. It was agreed that these should be freely available. However, individual scientists and institutes should evaluate whether it is worth adopting a new and complicated technique. Since the process of setting up complex protocols may well lead to the use of high numbers of animals, investigators should consider collaborating with colleagues who perform similar experiments at large scales.

Ethical considerations let us come to the understanding that there is an intrinsic value of life. We found that it is for this reason that it is morally wrong to kill more animals than absolutely necessary. Biomedical science is tasked with producing answers to pressing questions on the molecular functions of life and disease and finding new cures. It was pointed out that the principles of the 3R’s have to be respected at all times, but a number of animal experiments are indispensable. In this context, it is unavoidable to breed animals that are not used for these experiments, but it is important to ensure that their numbers are kept to a minimum.

Boris Jerchow
Member of ISTT’s Executive Council
October 23, 2013

List of participants and affiliations, excluding those who were unable to send permission for disclosure:

van der Broek, Frank, NVWA, The Netherlands; Aleström, Peter, The Norwegian Zebrafish Platform, Norway; Benavides, Fernando, University of Texas, USA*; Bussell, James, Wellcom Trust Sanger Institute, UK*; Chrobot, Nichola, MRC Harwell, UK; van Es, Johan, Hubrecht University, The Netherlands; Fentener van Vlissingen, Martje, Erasmus MC, The Netherlands; Hendriksen, Coenraad, InTraVacc, The Netherlands; Hohenstein, Peter, Roslin Intitute, UK*; Krimpenfort, Paul, NKI, The Netherlands; Morton, David, UK; Prins, Jan-Bas, LUMC, The Netherlands; Raspa, Marcello, EMMA, Italy*; Tramper, Ronno, Consultant, The Netherlands; van der Valk, Jan, NKCA; Wilbertz, Johannes, Karolinska Institutet, Sweden*; Ohl, Frauke, Utrecht University, The Netherlands; Pool, Chris, KNAW, The Netherlands; Witler, Lars, Max-Planck Institute Mol. Gen., Berlin, Germany*.

* ISTT members

Workshop: “Animals bred, but not used in experiments”, October 18-20, 2013, Hotel Duin & Kruidberg, Santpoort, the Netherlands (Picture kindly provided by Fernando Benavides)

Workshop: “Animals bred, but not used in experiments”, October 18-20, 2013, Hotel Duin & Kruidberg, Santpoort, the Netherlands (Picture kindly provided by Fernando Benavides)


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