Sdr9c7^em1Maak

A crRNA (5-CAACCAGTTGTTTGGCCAAC-3) targeting exon 1 and a trans-activating crRNA (tra-crRNA) were used with CRISPR/Cas9 technology to create a knockout allele. An ssODN (5-TCTTCATCACTGGCTGTGACTCCGGCTTTGGGAATCTGTTGGCCAAACAACTGGTTGATAGGGGCATGAAAGTGCTTGCT-3) was included to include a silent (synonymous) c.117C>T mutation to avoid cleavage by Cas9, reducing the chances of creating potentially embryonic lethal bi-allelic mutations. The resulting mutation is a single basepair deletion (c.123delG) in exon 1. Lack of protein expression from this allele was confirmed by immunohistochemistry. (J:295441)