NFKB2 Knockdown Reduces Expression of HR-Related Genes

Homologous recombination (HR) serves multiple roles in DNA repair that are essential for maintaining genomic stability. The central HR protein, RAD51, is frequently overexpressed in human malignancies, thereby elevating HR proficiency and promoting resistance to DNA-damaging therapies. Here we find that non-canonical NF-κB factors control the expression of RAD51 andother HR-promoting proteins. Transcriptional silencing of p100/p52 reduces RAD51 protein levels in multiple human cancer subtypes. RNA-seq after p100/p52 silencing identifies RAD51 as the most differentially expressed gene among 40 genes with known relevance to HR. While p100/p52 depletion inhibits HR function in human tumor cells, it does not influence the function of other double-strand DNA repair pathways including single-strand annealing, microhomology mediated annealing, or non-homologous end joining. U2OS cells were transfected with siRNA at 75 nM total using Lipofectamine RNAiMAX. Total RNA was collected 48 hours later and submitted to the Functional Genomics Facility core at the University of Chicago for QC, library preparation, and sequecing using an Illumina HiSeq 4000 instrument.