Figure 1: CRISPR RNP delivery.
HEK293T cells were transfected in triplicate with RNP containing CRISPRcraft™ S.p. Cas9 Nuclease and guide components from the CRISPRcraft™ S.p. Cas9 Nuclease Control Kit targeting HPRT, using either lipofection or electroporation. Transfection conditions: TransIT-X2® Dynamic Delivery System (Mirus), 50 nM gRNA and 25 nM Cas9; Lipofectamine® RNAiMAX (ThermoFisher Scientific), equimolar Cas9 and gRNA at 25 nM; Ingenio® Electroporation Solution (Mirus), 1.5 µM gRNA and 0.75 µM Cas9, plus Alt-R® Cas9 Electroporation Enhancer (IDT). At 48 hours post-transfection, cleavage efficiency at the HPRT locus was determined using the T7E1 mismatch detection assay and is expressed as % gene modification. |