CARHSP1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 near-haploid human cell line. This product provides a loss-of-function model for the calcium-regulated heat-stable protein 1 (CARHSP1) gene, enabling the study of CARHSP1-mediated post-transcriptional gene regulation. The polyclonal population contains a diverse array of knockout alleles generated by CRISPR/Cas9-mediated gene disruption, maintaining the cell line??s genetic tractability while avoiding the clonal selection that may introduce secondary mutations.
HAP1 is a fibroblast-like, adherent cell line derived from a male patient with chronic myeloid leukemia (CML). Its near-haploid karyotype makes it an exceptional model for genetic screens and haploid genetics, as the presence of a single allele for most genes simplifies functional genomics studies. HAP1 cells retain key signaling pathways relevant to cancer biology, including those driving proliferation and survival, providing a physiologically relevant context for studying gene function in leukemia and other malignancies.
CARHSP1 encodes a calcium-regulated RNA-binding protein that binds AU-rich elements (AREs) in target mRNAs, stabilizing them and enhancing translation. Its activity is regulated by phosphorylation via calcium/calmodulin-dependent kinases, MAPKs (ERK, p38), and protein kinase C. CARHSP1 interacts with calmodulin, 14-3-3 proteins, and competes with ARE-binding proteins HuR and TTP. It stabilizes mRNAs encoding TNF-??, IL-6, cyclin D1, and apoptosis-related transcripts, linking calcium signaling, MAPK cascades, and mRNA stability.
In HAP1 cells, the near-haploid background allows unambiguous assignment of phenotypic changes to CARHSP1 disruption, eliminating dominant effects that might mask loss-of-function phenotypes. Given the role of CARHSP1 in promoting cell survival and proliferation through ARE-mRNA stabilization, knockout HAP1 cells enable the dissection of post-transcriptional mechanisms underlying leukemogenesis and cancer progression. The CML origin of HAP1 provides a relevant context for investigating how altered RNA turnover contributes to hematopoietic malignancy, while the cell line??s adherent, fibroblast-like morphology facilitates standard cell-based assays.
CARHSP1 Knockout HAP1 Polyclonal Cells are ideal for post-transcriptional gene regulation studies, mRNA stability assays, and mapping RNA-protein interactions via RIP. These cells enable kinase inhibitor screening to evaluate MAPK pathway effects on ARE-mediated mRNA stabilization. They are compatible with haploid genetic screens to discover synthetic lethal interactions or modulators of CARHSP1 function. Typical assays include calcium flux measurements, phospho-specific western blotting, and flow cytometry for cell cycle or cytokine profiling. For further information, contact Ascent Research.
