C2CD5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the human C2CD5 gene within the near-haploid HAP1 cell line. This polyclonal pool provides a loss-of-function model suitable for investigating the functional roles of C2CD5 without the clonal selection steps required for monoclonal lines, thereby retaining the genetic diversity inherent to polyclonal populations. The product enables robust and reproducible knockout studies in a defined genetic background, facilitating efficient interrogation of C2CD5-dependent pathways.
The HAP1 host cell line is a human near-haploid adherent cell model originally derived from the KBM-7 chronic myeloid leukemia line, exhibiting fibroblast-like morphology. Its haploid karyotype simplifies genetic manipulation and phenotypic analysis, making it a preferred tool for functional genomics screens, gene-trap studies, and reverse genetics. HAP1 cells are widely used in both academic and industrial settings to dissect signaling pathways, validate drug targets, and conduct genome-wide loss-of-function screens, offering a streamlined system for exploring gene function in a human cell context.
C2CD5 is a C2 domain-containing protein that functions as a substrate for protein kinase A (PKA) downstream of GLP-1 receptor (GLP-1R)-mediated cAMP signaling. Upon GLP-1R activation, adenylyl cyclase generates cAMP, which activates PKA. PKA then phosphorylates C2CD5, enhancing its interaction with SNARE complex components??including syntaxin, SNAP-25, and VAMP??to facilitate insulin vesicle exocytosis. Additionally, C2CD5 exhibits calcium-dependent membrane binding, linking calcium signals to the exocytotic machinery. This positions C2CD5 at a critical convergence point between GLP-1R/cAMP/PKA and calcium signaling in the regulation of vesicle trafficking and secretion.
In the HAP1 cell model, disruption of C2CD5 provides a simplified yet informative context for studying conserved mechanisms of exocytosis and membrane trafficking. Although HAP1 is not a pancreatic ??-cell line, the core components of the exocytotic machinery??including SNARE proteins and calcium sensors??are broadly expressed. Therefore, C2CD5 knockout HAP1 cells serve as a tractable system to dissect C2CD5-mediated membrane fusion events, evaluate its role in vesicle dynamics, and screen for small molecules that modulate these processes. The haploid nature of HAP1 further enhances the utility of this model for genetic interaction studies and the discovery of novel regulators of the secretory pathway.
Researchers can employ these polyclonal knockout cells in diverse applications, including investigation of calcium-dependent vesicle trafficking, analysis of C2CD5 phosphorylation-dependent functions using western blotting and co-immunoprecipitation, and phenotypic assessment via immunofluorescence and calcium imaging. The cells are also compatible with functional assays such as pHluorin-based vesicle release measurements and RT-qPCR for transcript quantification. These capabilities support studies in diabetes research, metabolic syndrome, and drug screening for insulin secretion modulators. For additional technical information and ordering details, please contact Ascent Research.
