Fig. 5
From: Anti-CRISPR-mediated control of gene editing and synthetic circuits in eukaryotic cells
Cells with integrated AcrIIA4 become “write-protected” against future editing. a HEK293T cells were lentivirally transduced with a cassette constitutively expressing AcrIIA4. A clonal line was isolated and compared to untransduced wild-type cells. Cas9 + sgRNA was delivered either by a plasmid expressing both components or purified ribonucleoprotein complex (RNP). b Results from a T7E1 assay comparing editing efficiency between wild-type (WT) and AcrIIA4 (WPC) line targeting the PD1 locus run on an agarose gel. 100 bp standard is marked on left lane with positions of 500 and 1000 bp bands noted. Predicted lengths for uncut (996 bp) and cleaved products (437 and 559 bp) are annotated. Source Data are provided as a Source Data file. c Editing efficiency as quantified by TIDE analysis, with plasmid delivery, unsorted (plasmid) and sorted (sorted), as well as RNP delivery at various genomic loci. Error bars indicate s.e.m. of n = 2 experimental replicates for plasmid delivery and an estimate of technical variance (s.d.) of a single experimental replicate for RNP delivery. The dotted line is an estimate of detection sensitivity computed from sequencing traces of unedited cells (mean + 2× s.d.). Source Data are provided as a Source Data file
