Tmprss2^{em1(TMPRSS2)Synbl}

Cas9 and gRNA Tmprss2-crRNA_5 (protospacer sequence 5-GTGCTCAGACTGATACCCG-3) are used to introduce the following sequences at codon 19 in exon 3: GGSG linker, P2A, human TMPRSS2 cDNA coding sequence with SV40 intron inserted for enhanced expression, 1837 bp human TMPRSS2 3UTR, 112 bp human intergenic sequence immediately 3 of TMPRSS2 polyadenylation region , a cassette comprised of a mouse U6 promoter-driven Tmprss2-gRNA5 guide RNA with 2xPolIII terminator, all in reverse orientation relative to Tmprss2 gene, splice acceptor and SV40 polyadenylation sequence followed by a Flp Recombinase Target (FRT) site and 3-frame ATG short open reading frames. This cassette is designed to trap and truncate any mouse Tmprss2 isoforms that may splice past the hTMPRSS2 cassette. The FRT site allows Flp-mediated collapse of targeted tandem integration events if required. (J:101977)