Fig. 2.

Validation of individual BRCA1 variants inducing BRCA1 dysfunction. a Cell viability analysis of HAP1-BE3 cells transfected with each candidate gRNA using targeted deep sequencing. The gRNAs inducing c.4527C>T and c.3598C>T were used as a negative and a positive control, respectively. b Functional validation of BRCA1 variants using the CRISPR-based HDR method. c 5′-UTR reporter assays confirming the transcriptional repression of BRCA1 by the −97C>T mutation. d Cell viability analysis of the intronic mutations c.4986+3G>A and c.4986+5G>A induced by a single gRNA. Targetable C:G pairs and PAM are in red and underlined, respectively. Exon is shown as a rectangle. e In silico analysis of c.4986+3G>A and c.4986+5G>A using Human Splicing Finder, SpliceView, and NetGene2. Duplicate wells for each gRNA at each time point were processed. Error bars show the standard error of the mean