The CBARP Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CBARP gene. This loss-of-function model was generated through CRISPR/Cas9-mediated gene targeting in HAP1 cells, producing a heterogeneous collection of edited alleles. The polyclonal format mitigates clonal artifacts and provides a robust system for studying CBARP-dependent phenotypes, particularly in the context of calcium signaling, and for high-throughput screening of pharmacological modulators.
The HAP1 host cell line is a near-haploid human chronic myeloid leukemia model, originally derived from KBM-7 and characterized by the BCR-ABL fusion oncogene. Its haploid genome simplifies the creation of functional knockouts, as single-allele disruptions often suffice to eliminate gene expression without the need for biallelic editing. HAP1 cells maintain expression of key components of the calcium signaling pathway, making them a suitable platform for investigating voltage-gated calcium channel regulation and downstream effects.
CBARP functions as a modulator of voltage-gated calcium channels (VGCCs) by directly binding to the regulatory beta subunit CACNB4, influencing channel trafficking and gating. Its activity is regulated by cAMP/PKA signaling, membrane depolarization, and calcium influx, and it impacts downstream mediators calmodulin, CaMKII, and calcineurin. CBARP interacts with alpha subunits CACNA1A and CACNA1C to fine-tune calcium signaling in excitable cells, relevant to neuronal and cardiac function.
Disruption of CBARP in the HAP1 background creates a genetically tractable model to explore its role in VGCC regulation without confounding diploid compensation. The near-haploid state enhances the clarity of genotype-phenotype relationships and enables efficient knockout validation. This model is particularly suited to studying the molecular mechanisms underlying channelopathies, including neurological disorders and cardiac arrhythmias. Loss of CBARP allows researchers to examine alterations in calcium channel kinetics, secondary messenger activation, and calcium-dependent transcriptional programs.
Key applications of the CBARP Knockout HAP1 Polyclonal Cells include quantitative calcium imaging to track intracellular Ca2? dynamics, patch-clamp electrophysiology to measure VGCC currents, and co-immunoprecipitation to verify CBARP-CACNB4 interactions. The cells also facilitate drug screening for calcium channel modulators, CRISPR editing validation, and disease modeling. Typical assays include western blotting for CaMKII phosphorylation, RT-qPCR for gene expression changes, and immunofluorescence for channel localization. For further technical details, please contact Ascent Research.
