The CAPS2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of CAPS2 in a human embryonic kidney background. Generated by CRISPR/Cas9-mediated gene disruption, this polyclonal pool ablates CAPS2 expression to enable population-level investigation of calcium-dependent secretion. The model retains the fast growth and high transfection efficiency of HEK293T cells, making it ideal for functional analyses without clonal bias.
The HEK293T cell line originates from human embryonic kidney cells and stably expresses SV40 large T antigen, allowing episomal plasmid replication and exceptional transfection efficiency. Widely applied in protein expression, viral production, and signaling studies, these epithelial cells provide a robust platform for genetic manipulation. Their constitutive and inducible exocytotic machinery makes them suitable for studying vesicle priming and fusion proteins like CAPS2. The well-characterized epithelial morphology and rapid proliferation support reproducible experimental outcomes.
CAPS2 encodes a calcium-binding protein that senses intracellular Ca2+ and orchestrates dense-core vesicle exocytosis. Mechanistically, CAPS2 binds Ca2+, interacts with phosphatidylserine and SNARE proteins (syntaxin-1, SNAP-25, VAMP2), and stabilizes trans-SNARE complexes to drive vesicle fusion. Upstream regulators include Ca2+ influx, cAMP/PKA, CREB, and BDNF/TrkB signaling. CAPS2 cooperates with synaptotagmin in the synaptic vesicle cycle to couple calcium elevation to neuropeptide and hormone release.
CAPS2 disruption in HEK293T impairs calcium-triggered dense-core vesicle fusion, modeling secretion defects associated with autism spectrum disorder and neurodevelopmental conditions. The loss-of-function phenotype can be rescued by re-expressing CAPS2 variants for structure-function studies. The polyclonal population preserves cellular heterogeneity, reflecting tissue-level responses more closely than monoclonal lines. This model is valuable for dissecting SNARE-mediated exocytosis requirements and testing mutations linked to intellectual disability.
Research applications include Fluo-4 calcium imaging, co-immunoprecipitation of SNARE complexes, and ELISA-based secretion assays. Knockout validation is performed via immunofluorescence, western blotting, and RT-qPCR. The cells are suitable for screening exocytosis modulators and drug discovery for autism spectrum disorders. Additionally, functional complementation assays can identify critical domains within CAPS2 or interacting factors required for secretion. Contact Ascent Research for further details and custom lot requests.
