Per2^em1Vrshp

CRISPR/Cas9 genome editing was used to insert TCC AGT to TCA GCT mutations in codons 477 and 478, resulting in the insertion of a serine to alanine point mutation in codon 478 (S478A). TGuide RNA, donor oligonucleotide, and the Cas9 nuclease were introduced into the cytoplasm of embryos derived from a cross of C57BL/6J males and Per2tm1Jt-derived embryos with well recognized pronuclei. PCR amplicon sequencing was used to distinguish between the 129 and C57BL/6J alleles and founders with the S478A allele inserted in cis with Per2tm1Jt in the 129 allele were selected. This allele includes the S478A mutation and a luciferase gene and floxed neo cassette inserted between exon 23 and the 3' UTR. (J:288606)