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. 2020 Jan 27;21(3):818. doi: 10.3390/ijms21030818

Figure 1.

Figure 1

Mpst gene targeting in mice. (A) Outline for the Mpst gene deletion by CRISPR/Cas9 and production of 3 mutants. The 7304 bp mouse Mpst gene consists of 3 exons and is located proximal to its homolog Tst gene. The upstream (u) and downstream (d) crRNAs were designed to delete exon 2 which contains the start ATG codon and 67% of the entire open reading frame. Three independent mouse lines (1st, 2nd, and 3rd) were established. (B) Initial screening of 1st–3rd mouse lines from 14 independent mice (9 males and 5 females) that originated from individual fertilized zygotes electroporated with Cas9 protein, tracrRNA and crRNAs (u and d). PCR with forward (f) and reverse (r) primers detected the deletion of Mpst exon 2 in the 1st–3rd lines. (C) PCR detection of 1st and 3rd-type deletion using 1, 3, and r primers and 2nd-type deletion using 2, 4, and r primers from tail DNAs of wild-type (WT), Mpst-heterozygous (Het), and Mpst-homozygous (KO) mutant mice.