The CC2D1A Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the CC2D1A gene has been disrupted to create a loss-of-function model in the widely used HEK293T human embryonic kidney cell line. This pooled polyclonal format provides a heterogeneous mixture of gene-edited cells, enabling robust and reproducible functional studies without the clonal selection biases inherent in single-cell-derived lines. The targeted disruption of CC2D1A by CRISPR/Cas9 allows researchers to investigate the gene??s role in transcriptional regulation, neurodevelopmental signaling, and cancer biology within a high-expressing mammalian system.
HEK293T cells are a derivative of HEK293 cells that stably express the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication, leading to amplified protein expression and high-titer viral vector production. These human embryonic kidney epithelial cells are extensively utilized for transient transfection, recombinant protein production, and lentiviral packaging. The robust transfection efficiency and well-characterized signaling pathways make HEK293T an ideal host for studying the molecular mechanisms of CC2D1A without confounding endogenous expression from primary tissues.
CC2D1A encodes a calcium-binding transcriptional repressor that critically regulates neuronal development, cell proliferation, and differentiation. It directly binds the HTR1A promoter to suppress serotonin 5-HT1A receptor expression in a calcium-dependent manner. Additionally, CC2D1A modulates NF-??B and WNT/??-catenin pathways, interacting with the TSC1/TSC2 complex and the ESCRT-III subunit CHMP4B, thereby linking calcium signaling to endosomal trafficking and transcriptional control. Downstream targets include HTR1A, NF-??B components (p65/p50, I??B??), and WNT target genes. CC2D1A is activated by calcium ions and may be regulated by transcription factors such as Engrailed-2 (En2).
In the HEK293T background, CC2D1A knockout provides a tractable platform to dissect the gene’s role in pathways otherwise challenging to study in neurons or in vivo. High transfection efficiency permits rescue with wild-type or mutant CC2D1A to validate functional domains. The absence of endogenous serotonin receptors enables unambiguous HTR1A reporter assays. As a polyclonal population, it minimizes clonal artifacts, ensuring observed effects arise from uniform gene disruption.
These knockout cells are well-suited for a broad range of functional and biochemical studies, including HTR1A promoter-driven luciferase reporter and NF-??B reporter assays, RT-qPCR or Western blotting for downstream targets, co-immunoprecipitation of interaction partners such as Engrailed-2, TSC1/TSC2, and CHMP4B, immunofluorescence localization, and cell proliferation assays. The polyclonal knockout format ensures robust and reproducible results. Research areas include neurodevelopmental disorders, autism spectrum disorder, intellectual disability, and colorectal cancer susceptibility. For further technical assistance, please contact Ascent Research.
