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A loxP site flanked targeting vector containing a neomycin resistance gene under the control of the phosphoglycerate kinase, PGK, promoter was utilized in the construction of Mycntm1Psk. This selection cassette was inserted downstream of exon 3 of the targeted gene, and another loxP site was inserted upstream of exon 2. This construct was electroporated into 129 derived embryonic stem (ES) cells which were transiently transfected with a Cre recombinase vector to remove the selection cassette.
The Myctm2Fwa targeting vector was designed to place a loxP site between exons 1 and 2 and a loxP-flanked PGKneo cassette downstream of exon 3. The construct was electroporated into 129S6/SvEvTac-derived TC-1 embryonic stem (ES) cells. The neomycin cassette was removed by transient transfection with a Cre recombinase expressing plasmid leaving loxP sites flanking exons 2 and 3.
Varlakhanova N, Cotterman R, deVries W, Morgan J, Donahue LR, Murray S, Knowles B, Knoepfler P. myc maintains embryonic stem cell pluripotencaand self-renewal. Differentiation 2010; 80:9-19. (Medline PMID: 20537458).
ES Cell Sex and Genotype: These ES Cells are expected to be male (XY) and heterozygous/hemizygous for the mutation.
Requested cell lines are tested by PCR analysis for mycoplasma spp. and Murine Parvoviruses (MPV) upon completion of expansion and prior to shipment.
Mice derived from requested cell lines via Microinjection services will be pathogen tested prior to delivery of mice and health reports will be provided. If you require additional health status information prior to requesting microinjection please email csmmrrc@jax.org.
Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.
Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.