Pot1a^tm1.1Blas

An arginine to cysteine substitution at amino acid 117 was knocked-in using the FLEx switch system. A cassette containing the following elements replaced the wild-type exon 7: a loxP site, a loxP511 site, exon 7 with a mutation changing arginine codon 117 (CGC) to cysteine (TGC) (p.R117C) in reverse orientation, an FRT site flanked neomycin selection cassette, a loxP site in reverse orientation, wild-type exon 7 in forward orientation, and a loxP511 site in reverse orientation. The neomycin selection cassette was removed via Flp-mediated recombination. Upon expression of Cre recombinase, the wild-type exon 7 will be removed and the mutated exon, harboring the R117C missense mutation, will be flipped to the forward orientation. Sequencing transcripts from this allele indicates that the insertion of the mutant exon 7 disrupts proper processing of intron 6 as they retain the inverted mutant exon in that intron. (J:325969)