The GPS2 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells harboring targeted disruption of the GPS2 gene. This loss-of-function model enables investigation of GPS2-dependent transcriptional regulation and signaling networks without the need for clonal isolation. The polyclonal format preserves genetic heterogeneity while providing robust knockout efficiency across the cell pool, making it suitable for pooled functional screens and population-level analyses.
The parental HAP1 cell line is a near-haploid, adherent human cell line with fibroblast-like morphology, derived from the KBM-7 chronic myeloid leukemia (CML) background and of male origin. Its haploid state facilitates gene knockout and genetic screening, as only a single allele requires disruption to produce a null phenotype. HAP1 cells are widely employed in haploid genetic screens to assign gene function, validate drug targets, and model oncogenic processes within a myeloid leukemia context.
GPS2 functions as a transcriptional corepressor by integrating into the N-CoR-HDAC3 complex, which is recruited by unliganded nuclear receptors including RAR, TR, and PPAR?? to repress target genes. Additionally, GPS2 inhibits JNK MAPK signaling by stabilizing the phosphatase MKP7, thereby limiting JNK phosphorylation and downstream c-Jun activation. Through these mechanisms, GPS2 regulates p53 pathway activity and transcription of targets such as p21/CDKN1A, BAX, and IL6. GPS2 interacts with corepressors SMRT, TBL1, TBLR1, and UBC9, and is modulated by insulin and TNF??, positioning it at the nexus of metabolic, inflammatory, and genome stability pathways.
In the HAP1 CML background, disruption of GPS2 may alter JNK-mediated proliferation and apoptosis, as well as nuclear receptor-driven differentiation programs. This model is particularly valuable for dissecting the role of GPS2 in myeloid leukemia biology, where JNK signaling and p53 activity are frequently dysregulated. The near-haploid nature of HAP1 cells simplifies the interpretation of knockout effects and enhances the utility of the model for high-throughput chemical and genetic modifier screens.
Representative applications include functional genomics studies, drug target validation, and signaling pathway analysis in cancer, metabolic syndrome, and inflammation. Assays such as JNK phosphorylation Western blot, p53 reporter luciferase assays, and co-immunoprecipitation of GPS2 with N-CoR/HDAC3 can be performed. ChIP-qPCR can assess HDAC3 recruitment to target genes, and RNA-seq can reveal transcriptomic changes. The cells are also amenable to haploid genetic screens for synthetic lethal interactions or modulator identification. For more information, contact Ascent Research.
