Transgenic Core FAQs
General Questions
What does the Transgenic Core do?
We provide access to our micromanipoulation and embryos stem cell workstations along with necessary reagents:
- specialized plasmids
- embryonic stem (ES) cell lines
- FBS
- feeder cells certified for ES cell culture.
The Transgenic Core provides assisted reporductive technology for mice and rats including:
- in vitro fertilization to exand animal colonies or to prepare 2-cell eggs for cryoperservation
- cryropresrvation of 8-cell eggs
- sperm cryopreservation
- recovery of live animals from cryopreserved materials
- intracytoplasmid sperm insertion (ICSI)
The Transgenic Core derives specific pathogen free mice or rats from pathogen infected animals.
Hands-on training is proved to individuals in all aspects of transgenic technology.
Refer to the list of services.
How do I submit a transgene?
How much does it cost?
Members of the following University of Michigan centers receive discounts:
- Comprehensive Cancer Center
- Rheumatic Diseases Research Core Center
- Michigan Diabetes Research Center
- Michigan Gastrointestinal Peptide Research Center
- Michigan George M. O’Brien Renal Core Center
To submit a request for services please Login or Sign Up at our:
Plasmids and Cells Available with Material Transfer Agreements
When will my service request be processed?
The Transgenic Core prioritizes all requests for service on a “first-come, first-serve” basis. This standard is applied to everyone equally, even our faculty directors. Typically ES cell work is scheduled one to three months ahead. Since the inception of the Transgenic Core our policy has been that no project, whether it be gene targeting in ES cells or a transgenic mouse model, will enter the work queue until all of the required materials are provided. This includes both scientific materials, such as DNA samples and genotyping tests, and paperwork, such as approval to use animals in research, material transfer agreements and billing information.
How effective is the Transgenic Core?
What about consultation?
What kind of paperwork is involved?
- IACUC protocol approval number
- ULAM shortcode number for animal housing and veterinary care
How do I acknowledge Cores and the Rogel Cancer Center in research publications?
Referencing The CCSG:
When referencing the Core Grant in any of your publications or other materials, please use the following citation:
This research was supported (in part) by the National Institutes of Health through the University of Michigan’s Cancer Center Support Grant (P30 CA046592).
Referencing Shared Resources:
Please remember our Cores! When referencing any Cancer Center supported core in any of your publications or other materials, please use the following citation:
This research was supported (in part) by the National Institutes of Health through the University of Michigan’s Cancer Center Support Grant (P30 CA046592) by the use of the following Cancer Center Core(s): xxx
What is a transgenic mouse/rat?
What is involved in making transgenic mice/rats?
What can I do to maximize a successful transgenic outcome?
We guarantee that you will receive 3 or more transgenic mice or rats, however we can not guarantee transgene expression or transmission. The best strategy is to use a promoter that is already well characterize in transgenic mice or to employ very large flanking regions greater than 10 Kb. Alternatively, a bacterial artificial chromosome can be used to direct gene expression. BACs are include over 100Kb of genomic DNA sequence and often direct gene expression in a fashion which close matches the expression of endogenous genes.
The yield of transgenics is optimized by injecting highly purified linear DNA fragments with overhanging ends. Remove as much vector sequence as possible from the construct since prokaryotic sequences inhibit transgene expression. Although not a guarantee, demonstrated expression in a cell line is a positive indicator of in vivo expression and provides a rapid, inexpensive method to demonstrate that the transgene has been constructed properly. Contact Thom Saunders for more information on transgene design.
How many transgenic mice/rats will I get?
What if my DNA construct is lethal?
What is the significant of transgene copy number?
What if my transgene is too big?
Gene Targeted Mouse
What is a gene targeted mouse?
What is involved making gene targeted mice?
What can I do to maximize a successful gene targeting outcome?
- Characterize the genomic structure of your thoroughly.
- Develop a sensitive screen for homologous recombination in the ES cells.
- Consider the biological consequence of the mutation you introduce.
We can guarantee that we will inject your embryonic stem (ES) cells into a minimum of 50 blastocysts for ES cell-mouse chimera formation. Because of the intrinsic variability in individual ES cell clones, we can not guarantee that chimeras will be produced or that they will transmit your targeted gene through the germline. Therefore, we recommend that you provide at least three clones for microinjection. In collaboration with other labs on campus, we have successfully targeted over 60 genetic loci. We are confident in that we can work with you to genetically engineer new strains of mice that carry novel mutations of value to your research. The Core provides plasmids for gene targeting, ES cell lines that have been tested for germline chimera formation, feeder cells for ES cell culture, FBS tested for ES cell culture, and training in the exacting techniques required for successful ES cell culture. Contact Thom Saunders for more information on gene targeting projects.
NIH IRP Guidelines for the Availability of Transgenic/Knockout Animals
Transgenic and gene “knockout” animals that have been developed using NIH IRP (intramural research program) funds and resources will be provided to other laboratories following publication of descriptions of the animals in the peer reviewed literature. It is an obligation of NIH intramural scientists to make such animals widely available for research purposes. This can be achieved by making arrangements to send breeding pairs to a central repository such as the Induced Mutant Resource at the Jackson Laboratory. This would assure the availability of clean, genetically characterized animals within a year’s time. An attempt should be made to reduce duplication of effort by setting up collaborative experiments whenever possible; however, this should not be used as a mechanism to inhibit the distribution of animals.
These guidelines for the IRP are now in agreement with those of the US Public Health Service (PHS) for the extramural community: “It is the policy of PHS to make available to the public the results and accomplishments of the activities that it funds…Therefore, when these resources are developed with PHS funds and the associated research findings have been published or after they have been provided to the agencies under contract, it is important that they be made readily available for research purposes to qualified individuals within the scientific community. This policy applies to grants, cooperative agreements, and contracts.”
These guidelines supplement those already covered by the NIH Guide for other types of biological materials and resources. The NIH Guide is available online.
Review
Articles
Auerbach AB, Norinsky R, Ho W, Losos K, Guo Q, Chatterjee S, Joyner AL. 2003. Strain-dependent differences in the efficiency of transgenic mouse production. Transgenic Res. 12:59-69.
Adams DJ, van der Weyden L.Contemporary approaches for modifying the mouse genome. 2008. Physiol Genomics. 34::225-38.
Camper SA. 1987. Research applications of transgenic mice. Biotechniques 5, 638-650.
Capecchi MR. 1994. Targeted Gene Replacement. Sci. Am. March 1994. pp. 52-59.
Hughes ED, Qu YY, Genik SJ, Lyons RH, Pacheco CD, Lieberman AP, Samuelson LC, Nasonkin IO, Camper SA, Van Keuren ML, Saunders TL.2007. Genetic variation in C57BL/6 ES cell lines and genetic instability in the Bruce4 C57BL/6 ES cell line. Mamm Genome. 18:549-558.
Saunders TL. 2003. Reporter molecules in genetically engineered mice.Methods Mol Biol. 209:125-143.
Skarnes WC, Rosen B, West AP, Koutsourakis M, Bushell W, Iyer V, Mujica AO, Thomas M, Harrow J, Cox T, Jackson D, Severin J, Biggs P, Fu J, Nefedov M, de Jong PJ, Stewart AF, Bradley A. 2011. A conditional knockout resource for the genome-wide study of mouse gene function. Nature.474:337-342.
Books
Advanced Protocols for Animal Transgenesis: An ISTT Manual.2011. S Pease and TL Saunders (eds) Springer-Verlag, Berlin.
Culture of Animal Cells: A Manual of Basic Technique. Freshney, RI. 2005. Wliey-Liss. New York.
Shapiro Science – Book Stacks – 4th floor | QH 585.2 .F741 2005
Taubman Medical – Reference | QH 585.2 .F741 2005
Gene Targeting. Sedivy, JM, Joyner, AL. 1992. W.H. Freeman. New York .
Shapiro Science – Book Stacks – 4th floor | QH 442 .S431 1992
Gene Targeting: A Practical Approach, 2nd Edition. Joyner AL, ed. 2000. IRL Press at Oxford University Press. New York.
Taubman Medical QH 442 .G43851 2000
- Guide to Techniques in Mouse Development. Wassarman, PM, DePamphilis, MR, eds. Methods in Enzymology, vol. 225. Academic Press, New York.
- Shapiro Science- Book Stacks – QP 601 .C72
- Mammalian and Avian Transgenesis: New Approaches. Pease, S and Lois, C. 2005. Springer-Verlag. Berlin, Germany.
- Taubman Medical Library QH 442.2 .M2571 2006
Manipulating the Mouse Embryo: A Laboratory Manual. Hogan B, Beddington R, Constantini F, Lacy E. 1994. Cold Spring Harbor Press. New York.
Shapiro Science – Book Stacks – 4th floor | QL959 .M2651 1986Library Info
Manipulating the Mouse Embryo: A Laboratory Manual. Nagy, A, Gertsenstein, M, Vintersten, K, Behringer, R. 2003. Cold Spring Harbor Press. New York.
Shapiro Science – Book Stacks – 4th floor | QL 959 .M2651 2003
Methods in Enzymology. Guide to Techniques in Mouse Development. 1993. Edited by Paul M. Wassarman and Melvin L. DePamphilis. Vol. 225.
Taubman Medical Library –Call No: Journals
Mouse Genetics: Concepts and Applications. Silver, LM. 1995. Oxford University Press. New York.
Shapiro Science – Book Stacks – 4th floor | QH 432 .S561 1995
Teratocarcinomas and Embryonic Stem Cells; A Practical Approach.Robertson EJ, ed., IRL Press at Oxford University Press, 1987.
Transgenic Animals. Grosveld F, ed. 1992. Academic Press. New York.
Shapiro Science – Book Stacks – 4th floor | QH 442.6 .T73 1992
- Animal Transgenesis and Cloning. Houdebine, L-M. 2003. Wiley, John & Sons, Incorporated. Hoboken, New Jersey.
- Transgenic Mouse Methods and Protocols. Methods in Molecular Biology vol. 209. Hofker, MH,and van Deursen, J. 2002. Humana Press. Totowa, New Jersey.
- Shapiro Science – Book Stacks – 4th floor | QH 506 .M451 v.209
Transgenic Animal Technology: A Laboratory Handbook. 2nd Edition. Pinkert, CA. 2002. Academic Press, New York.
Shapiro Science – Book Stacks – 4th floor | QH 442.6 .T691 2002