Efficient Genome Editing with Cas9 mRNA and verified gRNA. HEK293T/17, U2OS and NHDF cells were co-transfected with 0.5 µg of Cas9 encoding mRNA, and 25nM verified gRNA using Turbo-mRNA Transfection Kit (0.5 µl/well of 24-well plate of both mRNA Reagent and Boost, OriGene Catalog TT100314). A T7E1 mismatch detection assay was used to measure cleavage efficiency at 48 hours post-transfection.