Fcgrt^{em1(FCGRT)Synbl}

Cas9 and gRNA Fcgrt-g91T (protospacer sequence 5- GGGTCACCCTGTCGGAATG-3) are used to introduce the following sequence at the start codon in exon 2: Kozak sequence and human FCGRT cDNA cassette inserted at the cleavage site, Sv40 intron, a 214 bp native hFCGRT 3UTR (3UTR), a 152 bp native human sequence 3 of the mapped 3UTR, and a mouse U6 promoter-driven guide RNA scaffold with 2xPolIII terminator (all in reverse orientation immediately 3 of the hFCGRT cassette), a splice-trap element with splice acceptor and SV40 polyadenylation sequence followed by 3-frame ATG short open reading frames. This active genetics cassette is designed to trap and truncate any mouse Fcgrt isoforms that may splice past the hFCGRT cassette. A Flp Recombinase Target (FRT) site is inserted to allow Flp-mediated collapse of targeted tandem integration events (if needed). (J:101977)