What does the Transgenic Facility do?
How do I request a service?
How much does it cost?
When will my service request be processed?
How effective is the Transgenic Core?
What about consultation?
What kind of paperwork is involved?
What is a transgenic mouse?
What can I do to maximize a successful transgenic outcome?
How many transgenic mice will I get?
What is the significant of transgene copy number?
What if my transgene is too big?
What does the Transgenic Facility do?
The Transgenic Facility routinely prepares transgenic mice and mice with targeted mutations for University of Kansas Medical Center investigators. These animals can be used to study gene function, expression, regulation, to develop animal models of human disease, to establish cell lines from specific cell types transformed in vivo, to produce mice with tissue-specific inducible gene expression or tissue-specific gene deletions, plus many, many more possibilities. Feel free to stop by with an idea and have our experience help guide you with your experiment.
We also derive specific pathogen free mice from pathogen infected mice. We provide hands-on training for individuals in transgenic technology. We offer cryopreservation of fertilized mouse eggs for investigators who wish to stop breeding a line and retrieval of cryopreserved stocks. Refer to the list of “Services and Pricing” in the navigation menu on our homepage and read the details of the service descriptions on the related links.
How do I request a service?
Follow the navigation menu on the left side of the home page, select “Services and Pricing”. After choosing the desired service, the submission form is at the bottom of the link.
How much does it cost?
University of Kansas Medical Center investigators can review the fee charges by going to the navigation menu on the left side of the home page and selecting “Services and Pricing”. Note that off campus investigators will pay an additional 50% fee of the listed price.
When will my service request be processed?
The Transgenic Facility prioritizes all requests for service on a "first-come, first-serve" basis. This standard is applied to everyone equally. The time between transgene DNA submission and microinjection is updated regularly; typically ES cell work is scheduled one week to one month after submission. Also, the Transgenic Facility policy has been that no project, whether it is 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?
We provide advice on all aspects of this technology from experimental design to mouse breeding. We can provide protocols and training for every step in the process of generating transgenic or gene targeted mice. We are ready to interact, our doors are open, please contact us with any questions by going to the navigation menu on the left side of the home page and selecting “Contact Us”.
What kind of paperwork is involved?
Any project that uses mice must be approved by the Institutional Animal Care and Use Committee (IACUC) and obtain space within the LAR for housing founder animals. In addition, an account number for billing and details about the DNA sample should be submitted prior to any work being done.
What is a transgenic mouse?
A transgenic mouse has a transgene in addition to its normal complement of genes. A transgene is an artificial gene cloned in the lab by recombinant DNA technology and microinjected into fertilized mouse or rat eggs. Eggs are transferred into foster mothers for gestation. Transgenic progeny are bred to produce a line. Transgenes integrate randomly into chromosomal DNA and are transmitted by Mendelian genetics.
What can I do to maximize a successful transgenic outcome?
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, such as a BAC or YAC construct. 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. Also, we can not guarantee transgene expression or transmission in transgenics.
How many transgenic mice will I get?
We guarantee that you will receive a minimum of two transgenic founders if we prep the DNA and genotype the pups, alternatively, if the investigator plans to prep the DNA and screen the transgenic mice, we guarantee 200 embryos will be injected and implanted with no guarantee on any founders produced. The purity of the microinjection DNA is the single most important factor which determines how many transgenic founders will be produced. Another very important parameter is the reliability and sensitivity of the screen for the transgene.
What is the significant of transgene copy number?
For the majority of transgenes the copy number does not correlate with expression level. The exceptions occur when large genomic fragments are used to make transgenics. Examples include P1 clones (90 Kb), BACs (180 Kb,) or YACs (400 Kb). Some reports suggest that if you use locus control regions or matrix attachment regions around your transgene that you may be able to insulate if form integration effects. Previous work in the literature has shown that attempts to produce low copy numbers by microinjecting dilute DNA does not affect copy number, but does reduce the overall yield of transgenic mice.
What if my transgene is too big?
The size of your transgene should not interfere with transgenic mouse production. Large transgenes may be difficult to clone. You may wish to consider BAC recombineering to produce large transgenes under the control of regulatory elements in the BAC. The facility has produced transgenics from Yeast Artificial Chromosomes up to 180 Kb in size. There are reports in the literature of transgenic mice produced from the microinjection of 90 Kb P1 clones, 248 Kb yeast artificial chromosomes, and even microdissected chromosome fragments.
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The University of Kansas Medical Center