The HPCAL1 Knockout HAP1 Polyclonal Cells provide a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the HPCAL1 gene in the HAP1 cell line. This loss-of-function model enables investigation of HPCAL1’s role in calcium and cAMP signaling without clonal selection, capturing a range of editing outcomes across the population. The polyclonal format is suited for pooled functional screens and studies requiring a diverse allelic series.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. With a haploid karyotype and BCR-ABL positivity, HAP1 cells serve as a robust platform for gene knockout experiments, benefiting from simplified genetic manipulation and reduced gene redundancy. Their hematopoietic origin and stable growth characteristics make them well-suited for signaling studies and high-throughput applications.
HPCAL1 encodes a neuronal calcium sensor protein that binds intracellular calcium via EF-hand domains and directly modulates adenylyl cyclase isoforms such as ADCY1 and ADCY8. This interaction regulates cAMP production, downstream PKA activity, and CREB phosphorylation, thereby linking calcium transients to transcriptional responses. HPCAL1 activity is triggered by upstream calcium elevation through GPCR-mediated release or neuronal depolarization, and it interacts with factors including neurocalcin delta and calmodulin. Within the broader network, HPCAL1 functions alongside CACNA1C and NCALD to coordinate calcium homeostasis and synaptic plasticity.
Although HAP1 cells are of hematopoietic origin and do not naturally express the full complement of neuronal signaling proteins, the HPCAL1 knockout model in this clean genetic background permits reductionist analysis of its calcium-dependent signaling mechanisms. Ectopic expression or agonist stimulation can reconstitute pathway components, allowing dissection of HPCAL1’s modulatory effects on adenylyl cyclase activity and cAMP dynamics independently of neuronal context. This system is particularly valuable for studying the core HPCAL1-ADCY axis without confounding neuronal-specific factors.
The HPCAL1 knockout polyclonal HAP1 cells are applicable to a range of functional assays, including cAMP ELISA to assess adenylyl cyclase activity, Fluo-4 calcium imaging to monitor calcium dynamics, and phospho-CREB immunofluorescence to evaluate downstream PKA/CREB pathway activation. They support drug target validation for neuropsychiatric disorders and genome-wide CRISPR screens aimed at identifying modifiers of HPCAL1-dependent signaling. Confirmatory analysis via western blotting, RT-qPCR, and Sanger sequencing ensures knockout verification. For further details or custom applications, please contact Ascent Research.
