The ATOH8 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ATOH8 gene in HEK293T cells. This loss-of-function model facilitates investigation of ATOH8-mediated processes without the need for single-cell cloning, retaining population heterogeneity that mirrors natural variation. The polyclonal format is particularly useful for studies where clonal artifacts are undesirable, such as in tumor biology and signal transduction research.
HEK293T cells are a widely used human embryonic kidney cell line derived from HEK293 cells through stable integration of the SV40 large T antigen. This enables efficient episomal replication of SV40 origin-containing plasmids, leading to high transgene expression and robust viral production. Their kidney epithelial origin and ease of transfection make them an ideal host for gene-editing experiments, particularly for genes implicated in renal development and disease.
ATOH8 encodes a basic helix-loop-helix transcription factor that acts downstream of Notch1, TGF-beta, BMP4, and Wnt3a signaling. Upon activation, it transcriptionally regulates target genes including CDH1, CDH2, SNAI1, TWIST1, CDKN1A, and CDKN1B, thereby controlling cell adhesion, migration, and proliferation. ATOH8 forms regulatory complexes with TCF3, TCF12, ID1, ID2, and EP300. Knockout of ATOH8 perturbs these interactions, disrupting downstream gene expression and potentially altering epithelial-mesenchymal transition (EMT) dynamics.
In the HEK293T background, ATOH8 disruption offers a tractable model to study how this transcription factor modulates TGF-beta/BMP and Notch-driven EMT programs. The SV40 large T antigen status may influence cell cycle regulation, providing a context to examine cross-talk between ATOH8 and proliferative signaling. This polyclonal knockout population allows assessment of heterogeneous responses, which is relevant for understanding variable EMT phenotypes in cancer.
Researchers can employ these cells to explore ATOH8 function in kidney development and congenital kidney diseases, model EMT in hepatocellular and colorectal carcinoma, and conduct high-throughput drug screens for Notch/TGF-beta pathway modulators. Representative assays include Western blotting and RT-qPCR for target gene validation, immunofluorescence for E-cadherin/N-cadherin localization, migration and invasion assays, Notch reporter assays, co-immunoprecipitation, RNA-seq, and ChIP-seq. For further inquiries, please contact Ascent Research.
