Extended Data Fig. 1: Construction of catalytically inactive CWCas12f (dCWCas12f) and confirmation of indel-null activity in HEK293T cells.
From: Hypercompact adenine base editors based on a Cas12f variant guided by engineered RNA
a. An agarose gel image showing cleavage pattern of plasmid vector by wild-type or several catalytically dead mutants of CWCas12f. Plasmid vectors were constructed to harbor protospacer and PAM sequence for sgRNA. Five microgram of plasmid vectors were incubated with 1 microgram of gRNA and 2 microgram of CWCas12f or dCWCas12f at 37 °C for 1 h. The cleaved vector samples were resolved on a 0.8% agarose gel. M denotes molecular ladders. The image corresponds to a representative experiment for three independent experiments. b. Indel efficiencies of CWCas12f or dead mutants of CWCas12f at an NLRC4 locus (5’-TTTAGAGGGAGACACAAGTTGATA-3’) in HEK293T cells. n = 3 independent experiments.
