Deletion of the Fpr1-3 gene cluster by two sgRNAs spanning 95 kb. ( a ) Schematic overview of the strategy to delete a 95 kb DNA fragment on chromosome 17. The exons Fpr1 and Fpr3 are labeled in blue and purple respectively, and the target sites are indicated by arrowheads. PAM sequences are in red following the target sequence highlighted in blue or purple. After deletion of the DNA fragment, the resulting genomic sequence is composed of the 5′ part of Fpr1 (blue) and the 3′ portion of Fpr3 (purple). The locations of PCR primers (F, Forward; R, reverse) are indicated by arrows. ( b ) (Top) Genotyping of the founders. PCR analysis of F0 mice injected with <t>Cas9</t> protein and sgRNAs listed in ( a ). Arrowheads indicate the founders with deletion of the 95 kb genomic DNA fragment. (Below) Sequencing data of the PCR products from two founders showing the joint Fpr1/Fpr3 genomic sequence. M, DNA marker. ( c ) Genotyping analysis of F1 progenies from two founders showing germ line transmission of the 95 kb deletion. M, DNA marker.
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