The BRCC3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with disruption of the BRCC3 gene. This heterogeneous knockout pool, containing a variety of mutant alleles, avoids clonal selection biases and provides a robust platform for studying BRCC3 function. The polyclonal format ensures versatility for large-scale screening and reproducible phenotypes in bulk assays.
HAP1 is a near-haploid human fibroblast-like cell line derived from KBM-7, widely used for genetic screens due to its single-copy genome. The adherent, epithelial-like cells exhibit rapid growth and are amenable to transfection, CRISPR editing, and high-content imaging, making them an ideal host for knockout studies.
BRCC3 is a Lys-63-specific deubiquitinase that functions in two complexes: the BRCA1-A complex in DNA repair and the BRISC complex in immune signaling. Within BRCA1-A, together with BRCA1, ABRAXAS1, RAP80, BRE, and NBA1, it localizes to DNA double-strand breaks to remove K63-linked ubiquitin from H2AX, modulating repair pathway choice. In the BRISC complex, BRCC3 deubiquitinates NLRP3 inflammasome components and the type I interferon receptor, regulating inflammatory responses. Upstream regulators include ATM kinase, TNF-??, and IL-1??; downstream effects involve ??H2AX foci, RIP1 ubiquitination, and NLRP3 activation.
The near-haploid HAP1 background, combined with polyclonal gene disruption, ensures a robust loss-of-function model, as most cells harbor inactivating mutations in the single BRCC3 allele. These polyclonal knockout cells are particularly suited for dissecting BRCC3??s role in DNA damage responses triggered by genotoxic agents and in inflammatory cascades induced by cytokines. The population-based approach captures diverse allelic effects, reflecting heterogeneous tumor cell behavior.
Applications include Western blotting for ??H2AX to assess DNA repair, comet assays for strand breaks, and drug sensitivity profiling with PARP inhibitors. For inflammatory studies, NLRP3 inflammasome activation can be assayed by IL-1?? ELISA, and interferon pathway activity via RT-qPCR of ISGs. This model is ideal for functional genomics, drug target validation, and pathway dissection. For more information, contact Ascent Research.
