Zfy1^em3Asah

Exon 4 was targed for modification using the CRISPR/Cas9 system. The sgRNA target sequence was 5'-GAAGCAGTCTTAGATTCCAGtgg-3' (PAM sequence in lower case). As this gRNA targets identical sequence in this gene and in Zfy2, the procedure yielded single knock-outs (one founder for each gene) as well as double knock-outs (10 founders). Genotyping of this knock-out allele found in double knock-out founder #2 revealed 6 and 4 bp insertions (2 bp apart) in the target sequence. (J:238696)