The BCAS3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the BCAS3 gene in the human embryonic kidney HEK293T cell line. This product comprises a heterogeneous mixture of cells harboring diverse gene disruptions, providing a loss-of-function model that avoids the artifacts and bottlenecks of monoclonal selection. The polyclonal format is advantageous for experiments requiring population-level analysis, including bulk biochemistry, gene expression profiling, and functional assays in which clonal variation could confound interpretation.
The HEK293T cell line is a widely utilized human embryonic kidney epithelial derivative that constitutively expresses the SV40 large T antigen. This feature facilitates episomal replication of plasmids containing the SV40 origin of replication, making HEK293T cells a preferred host for high-level transient protein expression and retrovirus packaging. The cells maintain a transformed phenotype, rapid growth, and ease of transfection, which are advantageous for gene perturbation studies. In the context of BCAS3 knockout, HEK293T provides a well-characterized platform to assess the gene??s role in cell proliferation, signaling, and cytoskeletal dynamics.
BCAS3 functions as a transcriptional coactivator for estrogen receptor alpha (ESR1) and potentiates NF-??B signaling, thereby driving the expression of proliferative and pro-migratory genes such as CCND1, BCL2, IL6, and MMP9. It is activated by estrogen (E2) via ESR1 and by tumor necrosis factor alpha (TNF-alpha) through the NF-??B pathway, with downstream effects on cell cycle progression and microtubule stability. BCAS3 physically interacts with ESR1, the NF-??B subunit RELA, histone deacetylase HDAC9, tumor suppressor TP53, and tubulin isoforms TUBA1A and TUBB, integrating hormonal, inflammatory, and cytoskeletal signals. Its association with HDAC9 and microtubules suggests roles in chromatin remodeling and intracellular trafficking, while the interplay with TP53 may link BCAS3 to survival pathways.
In HEK293T cells, which lack endogenous ESR1 expression, BCAS3 knockout provides a clean genetic background to study ESR1-independent functions, particularly NF-??B signaling and microtubule dynamics. Ectopic expression of ESR1 can reconstitute estrogen responsiveness, enabling comparative analysis of BCAS3??s coactivator function. The polyclonal pool is well-suited for proliferation, migration, and reporter assays, as well as for co-immunoprecipitation and immunofluorescence studies to map protein interactions and subcellular localization. This model thus aids in deconstructing the multiple signaling arms regulated by BCAS3 in cancer and beyond.
Applications include western blotting, RT-qPCR, MTT proliferation assays, transwell migration/invasion assays, NF-??B luciferase reporter assays, co-immunoprecipitation, immunofluorescence, and estrogen-responsive reporter assays. This model supports investigations into breast, ovarian, and prostate cancer mechanisms, drug target validation, and signaling crosstalk. For further information, please contact Ascent Research.
