The AR Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the androgen receptor (AR) gene in human HAP1 cells. This loss-of-function model abolishes AR-mediated transcriptional activity, providing a defined system for studying androgen signaling and AR-dependent cellular processes. The polyclonal format ensures a heterogeneous population with targeted gene disruption, suitable for pooled functional genomics and robust phenotypic analyses without clonal selection bias.
HAP1 is a near-haploid human chronic myeloid leukemia cell line derived from KBM-7 cells and engineered to express HPV16 E6/E7 proteins. This results in a stable near-haploid karyotype, making it an ideal host for functional genomics and CRISPR-based genetic screens, as disruption of a single allele yields unambiguous loss-of-function phenotypes. HAP1 cells are characterized by rapid growth and high transfection efficiency, supporting high-throughput applications.
AR is a ligand-activated transcription factor that mediates androgen signaling. Upon binding dihydrotestosterone (DHT), AR dissociates from HSP90 and HSP70, homodimerizes, and translocates to the nucleus, where it recruits coactivators (NCOA1, NCOA2, EP300) to AREs, activating targets like KLK3 (PSA), TMPRSS2, and FKBP5. AR activity is regulated by upstream factors including EGFR ligands, IGF-1, and kinases (MAPK1, AKT1), and intersects with PI3K/AKT, MAPK/ERK, and Wnt/??-catenin pathways. Interactions with coregulators (NCOA3, CREBBP, HDAC1/3) and transcription factors (FOXO1) modulate its function.
AR knockout in HAP1 cells eliminates androgen-dependent signaling, enabling dissection of AR-dependent and independent mechanisms. The near-haploid background enhances phenotypic penetrance, minimizing compensatory effects. This model is particularly valuable for prostate cancer research, where AR signaling drives proliferation and survival. The polyclonal population facilitates robust screening and reveals phenotypic consistency across diverse mutations.
Applications include functional genomic screens, drug target validation for androgen-independent prostate cancer, high-throughput screening of AR modulators using ARE-luciferase reporters, and investigation of non-genomic AR functions. Assays such as Western blotting, RT-qPCR for KLK3 and TMPRSS2, ChIP, immunofluorescence, and co-immunoprecipitation detail AR expression, target gene regulation, and protein interactions. Proliferation and drug sensitivity assays (e.g., enzalutamide) assess therapeutic responses. RNA-seq defines the androgen-responsive transcriptome. This knockout population also supports androgen insensitivity syndrome modeling. For further details, please contact Ascent Research.
