Supplementary Figure 9: In vitro cleavage assay testing the efficiency of the gRNA in the dsxFCRISPRh gene drive to cleave the dsx exon 5 target site with the SNP found in wild populations in Africa | Nature Biotechnology

Supplementary Figure 9: In vitro cleavage assay testing the efficiency of the gRNA in the dsxFCRISPRh gene drive to cleave the dsx exon 5 target site with the SNP found in wild populations in Africa

From: A CRISPR–Cas9 gene drive targeting doublesex causes complete population suppression in caged Anopheles gambiae mosquitoes

Supplementary Figure 9

An in vitro cleavage assay using an RNP complex of Cas9 enzyme and the gRNA used in this study was performed against linearised plasmids containing either wild-type (WT) target site in dsx exon 5 or the same site containing the single SNP found in wild caught populations (SNP). Products of the in vitro cleavage assay were purified and analysed on a gel. Both the WT and SNP-containing target sites are susceptible to the cleavage activity of the RNP complex as shown by the diminished high molecular band and the presence of the two cleavage products of the expected size. A dsx exon 5 target site containing the WT sequence complementary to the gRNA but without the PAM sequence was used as a control (‘no PAM’).

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