Flrt2^em1Jnk

CRISPR/cas9 methodologies are used to introduce an H170N (nucleotide change CAC to AAT, note AAC was originally reported) in exon 2. The mutation disrupts interactions between FLRT2 and its ligands UNC5C and UNC5D. A single guide RNA (sgRNA) was produced in which the PAM sequence overlaps the H170 codon (sequence 5' GCCCAACAGGCACGCTACTCagg; PAM site is denoted by lowercase letters). The PAM sequence is destroyed by the codon-altering mutation, thereby rendering correctly edited alleles immune to further cas9 cutting activity. (J:339867)