Figure 1. Generation of Leptin receptor knockout rat using CRISPR/Cas9 system.

(A) Target loci of Lepr were amplified using genomic DNA templates from founders. M: DNA molecular weight marker DL2000; WT: Template DNA was replaced with wild-type genomic DNA; 1-8: Founder rats generated by microinjection. (B) PCR products of the targeted fragment in the Lepr in rats were sequenced. The protospacer adjacent motif (PAM) sequence was underlined and highlighted in green; the targeting sites were red; the insertions were purple, lower case; insertions (+) or deletions (−) were shown to the right of each allele. The E3, E4 and E5 represents exon 3, exon 4 and exon 5 of Lepr respectively. (C) Protein level of LEPR in the liver tissues of WT littermates and Lepr^−/− rats were detected by western blot, using β-actin as normalization.