Gimap5^em1Khoe

CRISPR/Cas9 was used to insert LoxP sites flanking the second exon. Because the second exon contains the translation start site, deletion of this exon results in loss of the protein. The accuracy of the insert was confirmed through sequencing. The presence of the LoxP sites was confirmed through PCR genotyping using a 5 LoxP site PCR forward, 5-GATC GTGCAACGACATGGG-3; 5 LoxP site PCR reverse, 5-ACGACCTTGCCCGACTAGAG-3; 3 LoxP site PCR forward, 5-CCACCCAAGGAGAGTTACCC-3; and 3 LoxP site PCR reverse, 5-GTCAAAGACAAGCTTCCACAGT-3 primers. (J:260013)