Fig. 4: Programmable DNA ADP-ribosylation preferentially drives base mutagenesis in human cells lacking TARG1.
a, Reversion of ADP-ribosylation of ssDNA in human cells by the TARG1 protein. b, Experimental setup for introducing edits in the EMX1 gene in HEK293T cells using an oligonucleotide RT. ssODN, single-stranded oligodeoxynucleotide. c, Extent of templated recombination (top), indel formation (middle) or base mutagenesis (bottom) using EMX1 sgRNA1 in HEK293T cells with TARG1 intact (WT) or disrupted (ΔTARG1). Bars and error bars represent the mean and s.e.m. of three independent transient transfections without selection or sorting. d, Frequency of base substitutions across the sgRNA target in the absence of the oligonucleotide RT. Results are shown with DNA nicking by Cas9 intact (top) or disabled (bottom). e, Extent of base mutagenesis of the ADP-ribosylated thymine across 17 target sites in five genes. f, Experimental setup for editing in HEK293T cells using siRNAs to reduce TARG1 levels. g, Extent of base substitutions in HEK293T cells following siRNA-mediated silencing of TARG1 expression. In d, e and g, bars and error bars represent the mean and s.e.m. of three independent transient transfections without selection or sorting.
