The Mouse Resource Facility (MRF) stocks numerous mouse reagents, including genomic DNA, plasmids used for DNA constructs and hormones for superovulation for distribution to MD Anderson investigators. Additionally, the MRF maintains a small colony of Cre and lacZ transgenic mice and other reporter mice. These reagents are provided for a small fee to cover maintenance costs.
The MRF stocks a variety of plasmids commonly used in the construction of DNA for the manipulation of the mouse genome. These include plasmids containing the neomycin gene that confers G418 resistance, Cre-loxP containing plasmids, plasmids containing the FRT sites used by FLP recombinase, LacZ containing plasmids and various other useful plasmids.
The following plasmids may be obtained by the investigator for use in generating targeting constructs. Five-microliter (5-µl) aliquots of the plasmid of choice will be supplied to the investigator for bacterial transformation. As these are plasmids that were donated to the GEMF, the investigator should verify them by restriction digest and/or sequencing.
Contains the neomycin resistance gene expressed from the PGK promoter with the bovine growth hormone poly A site and flanked by lox P sites for easy removal. The vector used is pBluescript II KS (-). Directionality is from the T3 to T7 promoter (A).
Contains the neomycin resistance gene expressed from the PGK promoter with the bovine growth hormone poly A site and flanked by lox P sites for easy removal. The vector used is pBluescript II KS (-). Directionality is from the T7 to T3 promoter (B).
Contains a TK cassette flanked by polycloning sites. Obtained from Allen Bradley.
Contains the neomycin resistance gene expressed from the PGK promoter with the bovine growth hormone poly A site and flanked by frt sites for removal with flp recombinase.
IRES-Cre-pA FRT2 PGK-Neo-bpA
Contains a 2.4-kb XhoI/SmaI fragment with an IRES-Cre-pA gene obtained from plasmid IRES-Cre-pA, and a 2.5-kb EcoRV/BamHI fragment from plasmid FRT-PGK-Neo-bpA FRT-6 in pBluescript II KS (-). The neo gene is flanked by FRT sites for removal with flp recombinase.
Contains the hygromycin-resistance gene expressed from the PGK promoter. The gene is flanked by multicloning sites.
Contains a ~1.0-kb XhoI/BamHI fragment with the IRES-Cre and a ~1.4-kb BamHI/SalI fragment with the polyA site for Cre from plasmid pOG231 all in pBluescript II KS (-).
Contains a tau-LacZ gene which can be expressed from an IRES as well as a loxP-flanked neo gene expressed from the PGK promoter. Both the LacZ gene and the neo gene are flanked by polycloning sites.
Contains the puromycin-resistance gene expressed from the PGK promoter with the bovine growth hormone polyA site and flanked by lox P sites for easy removal.
A plasmid containing the Cre gene expressed from the CMV promoter for universal expression. Obtained from Steve O'Gorman.
A plasmid containing the flp-recombinase expressed from the CMV promoter that was manipulated to conform to the Kozak consensus sequence for higher levels of expression. Obtained from Francis Stewart.
Cost: There is no charge for plasmids
The MRF maintains a small number of Cre, GFP and LacZ-containing mice (some carried as frozen sperm), and will distribute these to MD Anderson researchers (only) upon request. To determine what mice are available, please contact Jan Parker-Thornburg.
Mice cannot be sent to external investigators. However, the GEMF will be happy to supply ordering information to external investigators.
Mice transgenic for the CMV-Cre gene. Cre is expressed in all cells constitutively. These mice are in a C57BL/6 background.
C57BL/6 transgenic mice containing the Cre recombinase gene expressed from the Zp3 promoter. "Cre is expressed exclusively in the growing oocyte prior to completion of the first meiotic division." (Lewandoski M, Montzka Wassarman K, Martin GR  Zp3-cre, a transgenic mouse line for the activation or inactivation of loxP-flanked target genes specifically in the female germ line. Current Biology 7, 148-151.)
A deletor strain of mice used for generalized high level Flp recombinase expression. The ROSA26- gene was targeted with the enhanced version of the site-specific recombinase FLP (Farley FW, Sorioano P, Steffen LS, Dymecki SM . Widespread recombinase expression using FLPeR (flipper) mice. genesis 28, 106-110.) Constitutive, high level expression of FLPe is seen in all cells. These mice are in a 129 background.
Mice that carry the green fluorescent protein driven by the chicken beta-actin promoter and the CMV intermediate early enhancer. "This strain can be used as a source of fluorescently marked cells or tissues. The transgene is expressed in all nucleated embryonic tissues...Though ubiquitous, expression levels vary between different organs" (The Jackson Laboratory Web site: Strain 003115). The reference for this strain is Hadjantonakis AK, Gertsenstein M, Ikawa M, Okabe M, Nagy A. (1998). Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mech. Dev. 76, 79-90. This line is maintained as a frozen sperm stock, and will require revival by in vitro fertilization at a cost of $1650.
Cost: There is a $39 charge per mouse. For excess stock animals (non-transgenic, retired breeders) there is a charge of $17 per mouse.
The GEMF can supply small amounts of chemicals for ES cell culture, embryo shipping and superovulation. We can also supply morulae for culture to blastocyst for both C57BL/6 and BL/6 albino strains. The cost varies for each chemical depending on the cost to generate it.
The GEMF can train limited numbers of people in pronuclear injection and ES cell culture. This will typically be a one-on-one training, and will require a substantial time commitment from the trainee. To avoid cross contamination, trainees cannot work with their own mice prior to coming in for daily training.
To obtain reagents, log in to https://mdanderson.ilabsolutions.com, register for an account, then go to the site to request service.