iMUT-seq analysis of over 20 different DSB repair siRNA mediated knockdowns or small molecule inhibitors

iMUT-seq profiles double-strand break (DSB) induced mutations at extremely high sensitive with single nucleotide resolution, allowing for the quantification of all mutation types, including chromosomal rearrangements, around endogenous DSBs. AID-DIvA cells are treated with or without 4-hydroxytamoxifen to induce DSBs, then with IAA to induce degradation of the DSB inducing AsiSI enzyme and are then incubated to allow the breaks to be repaired. The DSB loci are then PCR amplified from the genomic DNA and subjected to NGS to provide high depth mutation profiling. All samples are either untreated (m/0) or treated (p/4)with OHT and have three independent biological replicates, with the exception of ATRi which has 2 replicates and controlsi which has 6 replicates. This is because the siRNA were divided into two batches to create replicates 1-3 and 4-6 and each batch contained a controlsi sample for reference causing it to be in all 6 replicates.