BAP1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human near-haploid HAP1 cell line. This product constitutes a loss-of-function model in which CRISPR/Cas9-mediated gene disruption eliminates BAP1 protein expression, creating a heterogeneous pool of edited alleles. The polyclonal format avoids clonal selection biases, providing a robust system for population-level functional analyses.
HAP1 is a near-haploid human cell line originating from KBM-7 chronic myeloid leukemia cells, characterized by adherent fibroblast-like morphology. Its near-haploid karyotype simplifies genetic interventions and reduces gene dosage artifacts, making it ideal for knockout screens and isogenic cell model generation. The parental HAP1 line exhibits consistent growth and reproducible responses to DNA-damaging agents, ensuring reliable experimental comparisons.
BAP1 encodes a deubiquitinase that specifically removes monoubiquitin from histone H2A at lysine 119, a mark deposited by the Polycomb repressive complex to silence transcription. Through this activity, BAP1 de-represses gene expression and regulates DNA damage repair and chromatin remodeling. DNA damage activates BAP1 via ATM/ATR signaling, and BAP1 functionally interacts with protein partners including ASXL1, ASXL2, BRCA1, BARD1, FOXK1, FOXK2, HCF-1, OGT, and YY1. Downstream, BAP1 deubiquitinates targets such as HCF-1 and influences cell cycle regulators (e.g., CDKN2A) and apoptosis effectors. Thus, BAP1 integrates DNA damage signals with chromatin dynamics and cell survival decisions.
BAP1 knockout in the HAP1 background disrupts critical protein interaction networks that maintain genomic stability. The near-haploid state accentuates functional consequences of BAP1 loss, sensitizing cells to genotoxic stress and impairing apoptosis??phenocopying BAP1-deficient cancers like mesothelioma, uveal melanoma, and renal cell carcinoma. This model allows unambiguous interrogation of BAP1??s tumor suppressor mechanisms, particularly its coordination with BRCA1/BARD1 complexes and Polycomb-mediated repression, free from confounding paralog compensation.
These polyclonal knockout cells enable diverse applications in cancer biology and epigenetics, including tumor suppressor mechanism dissection, DNA damage response pathway analysis (e.g., ATM/ATR signaling), drug target validation, and chromatin regulation studies. Key experimental assays include Western blotting for BAP1 and H2AK119ub, RT-qPCR for target gene expression, ChIP-qPCR for histone modification mapping, co-immunoprecipitation of BAP1 interactors (e.g., ASXL1/2, HCF-1), apoptosis and cell cycle analyses, and drug sensitivity profiling with DNA-damaging agents. The polyclonal population format is especially suited for high-throughput screening and population-averaged biochemical readouts. For additional information and ordering, please contact Ascent Research.
