The CCDC47 Knockout HEK293T Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout population targeting the CCDC47 gene in HEK293T cells. This loss-of-function model enables interrogation of CCDC47-dependent processes without residual protein expression. The polyclonal format provides a heterogeneous pool of edited cells, capturing diverse CRISPR-induced disruptions and serving as a versatile base for clonal isolation or bulk functional studies. The product is supplied as ready-to-use cells under standard adherent culture conditions.
HEK293T is a widely used derivative of HEK293 cells, stably expressing the SV40 large T antigen to enhance episomal replication and high-efficiency transient transfection. These human embryonic kidney epithelial cells are prized for their robust growth, ease of manipulation, and capacity for high-level recombinant protein expression and lentiviral production. The CCDC47 knockout in this background provides a physiologically relevant platform for studying calcium signaling, ER homeostasis, and stress responses.
CCDC47 is an ER-resident calcium-binding protein that functions as a key co-factor for STIM1. Upon ER calcium depletion, CCDC47 promotes STIM1 oligomerization and puncta formation at ER-plasma membrane junctions, where STIM1 gates Orai1 channels to initiate store-operated calcium entry (SOCE). The resulting calcium influx activates calcineurin and NFAT transcriptional programs. Concurrently, CCDC47 modulates UPR signaling by influencing ER chaperones like calnexin and the expression of GRP78 and CHOP. Upstream regulators include ER calcium depletion and the transcription factor ATF4, placing CCDC47 at a nexus of calcium homeostasis and proteostatic stress pathways.
In HEK293T, CCDC47 knockout impairs STIM1 puncta assembly and SOCE, attenuating NFAT-driven transcription and calcium-dependent signaling. This deficiency may also sensitize cells to ER stress by perturbing UPR, offering a model for studying the interplay between calcium dysregulation and proteotoxicity. Linked to trichohepatoneurodevelopmental syndrome and cancer-associated calcium signaling, this polyclonal knockout enables disease mechanism exploration and high-throughput screening of CCDC47-STIM1-Orai1 modulators.
Key applications include thapsigargin-induced SOCE measurement via Fura-2 or Fluo-4 calcium imaging, western blot analysis of STIM1, Orai1, GRP78, and CHOP, and NFAT luciferase reporter assays. Co-immunoprecipitation verifies STIM1 interaction changes, immunofluorescence visualizes STIM1 puncta, and RT-qPCR profiles UPR targets. These tools support drug development targeting calcium homeostasis and ER stress pathways. For further information, contact Ascent Research.
