Figure 2 | Scientific Reports

Figure 2

From: CRISPR/Cas9-mediated deletion of a GA-repeat in human GPM6B leads to disruption of neural cell differentiation from NT2 cells

Figure 2

Targeting and deleting the GA-repeat at the GPM6B gene using eSpCas9(1.1). (a) Schematic overview of the designed strategy to target and delete the GA-repeat. Green lines: gRNA locations; black arrows: primer location; red arrows: restriction enzymes sites; red stars: introduced mutations at PAM sites; yellow double arrows: LHA (426 bp) and RHA (520 bp); and red rectangle: the deleted GA-repeat. (b) T7E1 assay confirmed the efficiency of the designed gRNAs, targeting the GA-repeat at GPM6B gene. The original gel image is presented in Supplementary Fig. Xa. (c) Sanger sequencing of the wild-type GPM6B and the edited C1 confirmed successful deletion of the GA-repeat. Yellow arrow, deletion site of GA-repeat; F, forward primer; R, reverse primer; LgRNA, left gRNA; RgRNA, right gRNA; LHA, left homology arm; RHA, right homology arm; L100, ladder 100 bp; C1, edited clone 1; WT, wild-type.

Back to article page