Archive for the ‘list’ Category

World Map of Transgenic Core Facilities

Saturday, April 7th, 2012
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World Map of Transgenic Core Facilities

World Map of Transgenic Core Facilities

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

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

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

World map of transgenic facilities

Monday, May 17th, 2010
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World Map and links to transgenic facilities from the ISTT web site

World Map and links to transgenic facilities from the ISTT web site

The International Society for Transgenic Technologies (ISTT) web site includes a list of transgenic laboratories and facilities located around the world. In addition, the web page now features a Google Maps script that indicates the geographical location of each of the transgenic facilities listed. If your facility is not yet included in this web page, please contact us (email: istt@transtechsociety.org) and we will include it right away. Thanks for your collaboration.

These black mice…

Wednesday, March 4th, 2009
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A C57 black mouse

A "C57 black mouse"

 All researchers, students or technicians, sooner or later, come across the same questions, namely: “-which exact strain are “these black mice” we have in the animal house?”, “- I’ve been told they were “C57 black” but, I realised there are MANY similar mouse strains under this generic name.”  Indeed, there are several “C57black”-related mouse strains, that are NOT genetically identical and, therefore, the exact knowledge and recognisition of the appropriate mouse strain you are working with (i.e. or the mouse strain from which the ES cell that you’re using was originally derived)  is crucial for the success (or fiasco) of the corresponding scientific project. This point was nicely addressed by Johannes Wilbertz, ISTT Vice-President, at the last TT2008 meeting, held in Toronto. Here, I’d like to briefly summarise the most common “C57 black” mice and provide the corresponding links for interested readers, to explore additional information.

C57black is the “common” name (argot) of the “C57BL/6″ mouse strain, that can be purchased and obtained from different animal providers which originally obtained their starting mouse colonies from distinct stocks ,or even from the same stock but in different times, thus resulting (by genetic drift) in genetically divergent mouse strains. Considering four of the major mouse providers (The Jackson Laboratory, Charles River, Harlan and Taconic) these are the different C57BL/6 mice they sell and distribute.

THE JACKSON LABORATORY

C57BL/6J    These are the original “/J” mice, JAX Mouse strain #000664.

C57BL/6NJ   These are the “original /N” mice, who came from Jax to NIH, where they were bred and established and eventually returned to Jackson Lab, constituting the JAX Mouse strain #005304.

In addition, there are other C57BL/6 – related mouse strains available from Jackson, including the albino C57BL/6J mice, called B6(Cg)-Tyr<c-2j>/J mice or JAX Mouse strain #000058.

CHARLES RIVER

C57BL/6J   (available outside US) These are in principal “equivalent” to the original C57BL/6J mice from The Jackson Laboratory, transferred to France in 1981 and to UK in 2004 and bred according to JAX protocols.

C57BL/6NCrl   These originated in The Jackson Laboratory, were transferred to NIH in 1951 and then to Charles River in 1974.

Thanks to a comment received from Kate Pritchett, from Charles River, it must be noted that, for UK-customers, there is currently a third additional C57BL/6 strain available from this animal provider, identified as “C57BL/6JCrl”, that will be eventually discontinued, due to the contract agreements in place between Charles River and The Jackson Laboratory.

HARLAN

C57BL/6JOlaHsd   (available in Europe) These were originated in The Jackson Laboratory and were transferred to OLAC in 1983.

C57BL/6JRccHsd   (available in Europe) These were originated in The Jackson Laboratory and were transferred through RCC in 1973.

C57BL/6NHsd   These were originated in The Jackson Laboratory and then were transferred to NIH in 1974 and from there to Harlan.

TACONIC

C57BL/6NTac   These were originated in The Jackson Laboratory and then were transferred to NIH in 1974 and from there to Taconic in 1991.

C57BL/6JBomTac   These were originated in The Jackson Laboratory and then were transferred to Hannover in 1971 and from there to Taconic in 1988.

The JM8.N4 and JM8.F6 ES cells, used in the EUCOMM-KOMP projects, have been derived from C57BL/6N mice. Note added: According to a comment posted by Tom Fielder (please, read below), these ES cells are thought to be derived (and therefore, chimeras obtained must be bred to) from a C57BL/6N mouse, without any additional information from the actual provider used at the time. C57BL/6NTac mice are used for breeding chimeras. Same mouse strain, C57BL/6NTac, was used to derive the VGB6 ES cells, used by Regeneron and the KOMP projects too.

KNOWN GENETIC DIFFERENCES BETWEEN C57BL/6J and C57BL/6N mouse strains

Not all C57BL/6J mice available from different providers are absolutely genetically identical. Likewise, not all C57BL/6N mice available from different providers are genetically identical. But, what is more important is that C57BL/6J animals (the mouse strain used for the generation of many knock-out mouse models, reported and described and cryopreserved in repositoires world-wide) and C57BL/6N animals (the mouse strain choosen by the International knock-out projects) are DIFFERENT. This must be taken into account if you plan to breed new KO mice, generated from targeted ES cells provided by EUCOMM or KOMP projects, with former KO mice, most likely generated using c57BL/6J-derived ES cells.

GENE MUTATIONS

C57BL/6J mice carry a mutation in the nicotinamide nucleotide transhydrogenase (Nnt) gene, whereas C57BL/6N mice carry the wild-type allele for this locus. Exceptionally, the substrain C57BL/6JBomTac carry also the wild-type allele for the Nnt locus, suggesting that this mutation occurred in The Jackson Laboratory after 1971, when mice were sent to Hannover.

The C57BL/6JOlaHsd mice carry a mutation in the alpha-synuclein (Snca) gene, whereas, appearently, the rest of C57BL/6 mouse strains carry the wild-type allele for this locus.

As reported by Jennifer Moran, through the MGI-List, 19 SNPs were initially identified to distinguish the C57BL/6J and C57BL/6N mouse strains, with additional ~200 SNPs available to discriminate the various C57BL/6 mouse strains.

Eventually, we all hope these and other SNPs will become available through common mouse genetic databases to be used universally.

Collection of videos of animal transgenesis at the ISTT

Thursday, February 26th, 2009
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58 videos availbale at the ISTT

58 videos available at the ISTT

Currently, we have 58 videos on different aspects of animal transgenesis, kindly and generously shared by various ISTT Members. Videos are available within the members-only area of the ISTT web site. If you are not yet ISTT Member, please consider registering to our Society.

The current list of ISTT videos include:

1) Pronuclear microinjection of DNA into a sheep fertilised oocyte

2) Microinjection of murine ES cells into a mouse blastocyst

3) Nuclear transfer in mouse embryos. Nucleus of a blastomer into a enucleated metaphase II oocyte

4) Intracytoplasmic Sperm Injection (ICSI). Handling thawn (dead) sperm heads incubated with DNA solution (collecting heads and preparing for microinjection) as reported in Moreira et al. 2006

5) Intracytoplasmic Sperm Injection (ICSI) Microinjecting (piezo) sperm heads into metaphase II oocytes as reported in Moreira et al. 2006

6) Pronuclear microinjection of DNA into a mouse fertilised oocyte

7) Pronuclear microinjection of DNA into a mouse fertilised oocyte II

8 ) Pronuclear microinjection of DNA into a mouse fertilised oocyte III

9) Pronuclear microinjection of DNA into a mouse fertilised oocyte IV

10) Pronuclear microinjection of DNA into a mouse fertilised oocyte V

11) Pronuclear microinjection of DNA into a mouse fertilised oocyte VI

12) Pronuclear microinjection of DNA into a mouse fertilised oocyte VII

13) Pronuclear microinjection of DNA into a mouse fertilised oocyte VIII

14) Pronuclear microinjection of DNA into a mouse fertilised oocyte IX

15) Pronuclear microinjection of DNA into a mouse fertilised oocyte X

16) ES cell microinjection into a mouse blastocyst I

17) ES cell microinjection into a mouse blastocyst II

18) ES cell microinjection into a mouse blastocyst III

19) ES cell microinjection into a mouse blastocyst IV

20) Picking up mouse ES cells into a microinjection pipette

21) Loading a pipette with mouse embryos

22) Oviduct (infundibulum) transfer of mouse embryos

23) Intracytoplasmic Sperm Injection (ICSI) Microinjecting (piezo) sperm heads into metaphase II oocytes

24) Removing tails from sperm heads with a piezo

25) Fill – Lentivrus injection procedure (loading lentivirus and cellular debris particles into  the pipette)

26) Subzonal Injection. Lentivrus injection procedure (releasing lentivirus and cellular debris particles in the perizonal space, under the zona pellucida)

27) Discharge – Lentivirus injection (releasing lentivirus and cellular debris particles out of the pipette)

28) 2-cell mouse embryos post IVF

29) How to grab a mouse with one hand? I

30) How to grab a mouse with one hand? II

31) How to grab a mouse with one hand? III

32) Mouse intraperitoneal (IP) injection

33) Application of ear tags to mice for identification purposes I (usually done at weaning)

34) Application of ear tags to mice for identification purposes II (usually done at weaning)

35) Mouse tail biopsy using a home-made mouse restraint device and a sharp red-hot blade attached to a soldering tool for cutting and wound cauterisation purposes I (usually done at weaning)

36) Mouse tail biopsy using a home-made mouse restraint device and a sharp red-hot blade attached to a soldering tool for cutting and wound cauterisation purposes II (usually done at weaning)

37) Mouse euthanasia by cervical dislocation I

38) Mouse euthanasia by cervical dislocation II

39) Visual phenotyping of spatial acuity in mice with the optomotor test as described in Lavado et al. 2006

40) Visual phenotyping of spatial acuity in mice with the optomotor test. Cylinder turns left, albino mouse turns random because it cannot see, as described in Lavado et al. 2006.

41) Visual phenotyping of spatial acuity in mice with the optomotor test. Cylinder turns right, albino mouse turns random because it cannot see, as described in Lavado et al. 2006

42) Visual phenotyping of spatial acuity in mice with the optomotor test. Cylinder turns left, pigmented mouse turns left too because it can seem as described in Lavado et al. 2006

43) Visual phenotyping of spatial acuity in mice with the optomotor test. Cylinder turns right  pigmented mouse turns right too because it can see, as described in Lavado et al. 2006

44) Mouse ICSI using the XYClone Laser

45) ES-cell injection into mouse 8-cell embryos using the XYClone Laser

46) ES-cell injection into mouse blastocysts using the XYClone Laser

47) Mouse ICM xenogenic-free excission using the XYClone Laser

48) Bovine embryo biopsy using the XYClone Laser

49) Bovine ICSI using the XYClone Laser

50) Mouse vasectomy (abdominal)

51) Oviduct transfer upstream of the ampulla. (Oviduct “hole in the wall” transfer technique) Transfer between the infundibulum and the ampulla

52) Laser-assisted ES cell injection into a mouse 8 cell embryo using the OCTAX Laser Shot system (MTG)

53) Mouse vasectomy (scrotal)

54) Non-Surgical Embryo Transfer Device NSET (ParaTechs)

55) Flushing a mouse oviduct

56) Washing fertilised oocytes after IVF (transferring from HTF to KSOM)

57) Stained mid-gestation lacZ reporter mouse embryos (Myf5, Kit, Pax3 and Dct lacZ reporter mouse embryos are shown in this order, audio available)

58) ES cell injection into mouse 8-cell embryos

“Transgenic list” (tg-l) is now the official list of ISTT

Wednesday, April 23rd, 2008
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transgenic-list (tg-l)

ISTT

 The transgenic-list (tg-l), the reference channel of discussions on scientific and technical issues related to the generation and analysis of transgenic animals and transgenic research in general, established in its current format since July 1996 by Peter Sobieszczuk, has become the official list of the International Society for Transgenic Technologies (ISTT).

To illustrate this agreement, established between Peter Sobieszczuk (as the tg-list owner and therefore responsible person of the list) and Lluis Montoliu (as President of the ISTT) the ISTT will recommend and promote the use of tg-list to discuss any topic related with transgenic research. Likewise, the tg-list will announce this agreement in all messages, with a link to the ISTT WEB site.

 

Patents in ES cell work

Wednesday, April 16th, 2008
LinkedIn

mouse ES cells

Recently, at the tg-list, it was discussed the issue of patents that apply in the standard ES cell work. Those patents include the usual positive-negative selection and the use of isogenic DNA. Of course, there are many more with modifications or alternative methodologies but I believe these two methodology are still the basic ones. 

According to the United States Patent and Trademark Office, these are the patents alive and active:

Positive-negative selection methods and vectors
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 5,464,764
Issued: November 7, 1995
Filed: February 4, 1993
Assignee: University of Utah Research Foundation (Salt Lake City, UT)

Cells and non-human organisms containing predetermined genomic modifications and positive-negative selection methods and vectors for making same
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 5,487,992
Issued: January 30, 1996
Filed: June 28, 1993
Assignee: University of Utah Research Foundation (Salt Lake City, UT)

Cells and non-human organisms containing predetermined genomic modifications and positive-negative selection methods and vectors for making same
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 5,627,059
Issued: May 6, 1997
Filed: June 5, 1995
Assignee: University of Utah (Salt Lake City, UT)

Cells and non-human organisms containing predetermined genomic modifications and positive-negative selection methods and vectors for making same
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 5,631,153
Issued: May 20, 1997
Filed: June 5, 1995
Assignee: University of Utah (Salt Lake City, UT)

Cells and non-human organisms containing predetermined genomic modifications and positive-negative selection methods and vectors for making same
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 6,204,061
Issued: March 20, 2001
Filed: January 9, 1997
Assignee: University of Utah Research Foundation (Salt Lake City, UT)

Cells and non-human organisms containing predetermined genomic modifications and positive-negative selection methods and vectors for making same
Capecchi; Mario R., Thomas; Kirk R.
US Patent: 6,204,061
Issued: February 10, 2004
Filed: November 28, 2000
Assignee: University of Utah Research Foundation (Salt Lake City, UT)

Gene targeting in animal cells using isogenic DNA constructs
Berns; Anton, Robanus Maandag; Els, te Riele; Hein
US Patent: 5,789,215
Issued: August 4, 1998
Filed: August 7, 1997
Assignee: GenPharm International (San Jose, CA)

High efficiency gene targeting in mouse embryonic stem cells
Berns; Anton, Robanus Maandag; Els, te Riele; Hein
US Patent: 6,653,113
Issued: November 25, 2003
Filed: February 19, 1999
Assignee: Genpharm International, Inc. (Mountain View, CA)

 

Interesting e-mail lists in the animal transgenesis field

Tuesday, February 12th, 2008
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E-mail @ Lists

There are plenty of e-mail lists that distribute information among their associates. For instance, at the private level, we have at the ISTT the istt_list that is mostly used to distribute society-related information among ISTT members.

At the public level, and useful for those working in the field of mouse or animal transgenesis, there are several e-mail lists that are worth joining, to stay in touch and up-to-date within our field. I will be listing here four of them. I am sure there are other e-mail lists that could be added later on through additional comments to this original post.

Transgenic-List (tg-list)

The Transgenic List is probably the best known and most frequently used e-mail list on animal transgenesis, and has been and still is widely and extensively used for all those working or interested in the field. Peter Sobieszczuk, the founder of tg-list, takes very good care of it. This list is held and offered from the Department of Life Sciences at the Imperial College in London, UK. If you are working with transgenic animals are are not yet a member of it you should strongly consider joining. According to its main WEB site the tg-list: “Established in its present form in July 1996, it includes amongst its members active researchers in transgenesis from the novice to experts in the field. Major keywords cover wide spectrum of disciplines discussed: homologous recombination, targeted mutagenesis, inducible expression, ES cells, microinjection, mouse genetics, animal husbandry.”

Mouse Genome Informatics-List (mgi-list)

The mgi-list is one of the e-mail lists offered by The Jackson Laboratory, through its extraordinary and extremely useful site on Mouse Genome Informatics (MGI). The mgi-list is a forum for the discussion of topics in mouse genetics, and also is being used to distribute MGI news and updates. Often, most topics on animal transgenesis are simultaneously discussed on the tg-list and the mgi-list.

Phenome-List

The Phenome-list is yet another useful e-mail list offered by The Jackson Laboratory through MGI. The phenome-list is a forum for strain characterization and phenotypic data collection. Recently, the phenome-list has been announced that it will be moving to another host (Google Groups), and, from now on, will be found through this WEB site.

COMPMED-List

The COMPMED-list is one of the e-mail lists held at and promoted by the American Association for Laboratory Animal Science (AALAS) and it has a wider scope. According to its description at its main WEB site: “CompMed™ is an e-mail list for discussion of comparative medicine, laboratory animals, and topics related to biomedical research. CompMed serves professionals working in these fields by providing an interactive, easy-to-use environment to share information and discuss topics of common interest. CompMed is limited to participants who are involved in some aspect of biomedical research or veterinary medicine, including veterinarians, technicians, animal facility managers, researchers, and graduate/veterinary students.”


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