Figure 2.

Genome Editing by Multiplexed VEsiCas

(A) Gene deletion using VEsiCas. Shown is gel electrophoresis analysis of the EGFP locus deletion obtained in HEK293-EGFP cells through Multi-VEsiCas (right) treatment or by transfection with SpCas9 together with specific sgRNAs (sgEGFP5 and sgEGFPBi) (left). The amount of deletion (percent) was quantified by densitometry. Arrowheads indicate the expected band corresponding to PCR amplification of the deleted EGFP locus. (B) Activity of VEsiCas delivering SpCas9 nickase. Shown are percentages of EGFP knockout cells following incubation with VEsiCas-n, carrying SpCas9 nickase loaded with sgRNA toward two sites in the EGFP locus (sgEGFP3gW and sgEGFPBi) or with sgCtr. NT, not treated. Data are presented as mean ± SEM for n = 2 independent experiments.