Determining the role of R-loops in transcription-associated mutagenesis

DNA mutations are the leading cause of cancer, arising from endogenous and exogenous sources. My research focuses on transcription-associated mutagenesis (TAM) of human DNA, an understudied cancer etiology. Our lab hypothesizes that R-loops (RNA/DNA hybrids) affect the occurrence of transcription-associated large insertion and deletion mutations by contributing to double-strand break formation. To test this hypothesis, I am generating knockouts of SETX and XPF, two genes involved in R-loop processing. By characterizing these knockouts, I will determine if TAM mutation rates increase with elevated R-loop accumulation and decrease with the loss of XPF-mediated R-loop processing as predicted.