Comparison of functional scores for BRCA1 variants assessed in multiplexed and singleton HDR.

A. Eighteen amino-terminal BRCA1 variants were tested in the singleton HDR assay for DNA repair function. In the first bar, the control siRNA was transfected into the cells, and in all the other samples the siRNA specific to the 3’UTR of BRCA1 was transfected. The encoded BRCA1 variant used to rescue the depletion of the endogenous BRCA1 was indicated. B. The functional assessments of 44 BRCA1 variants (18 from panel A and 26 published) was compared in both the HDR multiplexed assay (x-axis) and the HDR singleton assay (y-axis). The dotted lines indicate the functional cutoffs for the singleton assay, with functionally normal defined as a score of > -0.51 and non-function defined as 2 transformed. Discordant data points are colored in red. C. The functional scores from the current multiplexed HDR analysis (x-axis) were compared to the singleton scores provided from the ASHRA assay (y-axis) [36]. The functional scores for the singleton assay were log2 transformed. D. The functional scores from the multiplexed HDR analysis (x-axis) were compared to the singleton scores for Olaparib resistance [36]. The functional scores for the singleton assay were log2 transformed.