A targeting vector was designed to introduce the R155H mutation of the valosin containing protein gene (Vcp), commonly found in patients with Inclusion Body Myopathy with Early-Onset Paget Disease and Frontotemporal Dementia (IBMPFD). The targeting vector also contained a frt-flanked neo cassette downstream of exon 5. This construct was electroporated into 129/SvEv-derived embryonic stem (ES) cell line. Correctly targeted ES cells were injected into 129/SvEv blastocysts and the resulting chimeric males were bred to 129/SvEv females. The donating investigator reported that these VcpR155H mice were backcrossed to C57BL/6JEiJ mice for at least six generations prior to sending to The Jackson Laboratory Repository in 2013 (see SNP note below). Upon arrival, sperm was cryopreserved. To generate our living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6JEiJ female mice (Stock No. 000924).
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Four markers throughout the genome suggested an in complete backcross.
