The BAG6 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the BAG6 gene in the HAP1 human cell line. This polyclonal pool contains a heterogeneous mixture of loss-of-function alleles, providing a robust loss-of-function model without clonal selection. The product enables functional interrogation of BAG6 in a near-haploid genomic context, facilitating high-confidence genetic studies.
HAP1 cells are a near-haploid, fibroblast-like cell line derived from the KBM-7 chronic myelogenous leukemia (CML) line of male origin. Due to their haploid nature for most chromosomes, HAP1 cells are an ideal host for CRISPR-based functional genomics and genetic screens, as they eliminate the complication of a second allele, ensuring unambiguous genotype-phenotype relationships. This background is widely used for pathway dissection and drug target validation.
BAG6 (BCL2-associated athanogene 6) is a multifunctional chaperone integral to protein quality control, apoptosis regulation, and immune signaling. It acts in concert with HSPA8/HSC70 to triage misfolded proteins toward the proteasome or autophagy, and it interacts with the TRC40/GET pathway components for tail-anchored protein insertion. BAG6 is also known to regulate caspase-3 activation and to modulate NK cell ligand surface expression. Its activity is influenced by DNA damage signals via ATM/ATR and cellular stress, and it engages with RNF126 ubiquitin ligase and p300/CBP cofactors. Disruption of BAG6 impairs protein homeostasis, sensitizes cells to apoptotic stimuli, and alters immune cell recognition.
In the HAP1 cellular context, BAG6 knockout provides a clean system to dissect its roles in protein homeostasis and immune evasion. The near-haploid background minimizes masking effects from wild-type alleles, enabling clear assessment of BAG6-dependent phenotypes such as proteotoxic stress responses, caspase-3-mediated apoptosis dynamics, and changes in NK cell ligand presentation. This model is particularly valuable for cancer biology research, where BAG6-mediated immune modulation and protein degradation pathways are often dysregulated.
These polyclonal knockout cells are suitable for a range of downstream applications, including Western blotting to confirm BAG6 loss, apoptosis assays (e.g., Annexin V staining), proteasome activity measurements, co-immunoprecipitation of interacting partners like HSPA8, flow cytometric analysis of cell surface markers, and transcriptomic profiling via RNA-seq. By providing a heterogenous knockout population, the product enables robust functional studies in areas such as protein homeostasis, cancer biology, apoptosis mechanisms, and immune evasion. For detailed product specifications and ordering, contact Ascent Research.
