BHLHE40 Knouckout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for the transcriptional repressor BHLHE40 in an intestinal epithelial context. The polyclonal cell pool harbors heterogeneous CRISPR/Cas9-mediated gene disruptions, enabling robust functional studies without clonal selection artifacts.
HT29 is an adherent epithelial cell line isolated from a primary colon adenocarcinoma of a 44-year-old Caucasian female. These cells are widely employed as a model for colorectal carcinoma and exhibit characteristics of differentiated enterocytes, including polarisation and microvilli. Their well-characterized signaling networks and tumorigenic properties make them ideal for studying cancer-related pathways.
BHLHE40 encodes a basic helix-loop-helix transcriptional repressor that integrates signals from TGF-??, HIF-1??, and circadian clock pathways. It is activated by upstream regulators TGF-??1, HIF-1??, IL-6, p53, and Notch, and transcriptionally represses downstream targets CCND1, MMP1, BCL2, CCNB1, and IL-2. BHLHE40 interacts with BHLHE41, ARNT, BMAL1, CLOCK, and HDAC1 to form repressive complexes. Through these interactions, BHLHE40 modulates cell cycle progression, apoptosis, and epithelial-mesenchymal transition. In colorectal cancer, it acts as a tumor suppressor by repressing cyclin D1 (CCND1) and MMP1 expression, thereby inhibiting proliferation and invasion. The signaling network involves TGFBR1, SMAD2/3, HIF1A, ARNT, and TP53, positioning BHLHE40 at a convergence point of tumor-suppressive and circadian pathways.
In the HT29 colorectal adenocarcinoma model, BHLHE40 disruption may relieve repression of cell cycle and invasion genes, providing a system to study its tumor-suppressive functions. This knockout enables dissection of BHLHE40-mediated TGF-??-induced apoptosis, hypoxia-responsive gene regulation, and circadian rhythm effects on cancer cell behavior.
Typical experimental approaches include western blotting and RT-qPCR for expression analysis, RNA-seq for transcriptome profiling, ChIP-qPCR for assessing BHLHE40 binding to target promoters, flow cytometry for apoptosis and cell cycle analysis, migration and invasion assays to evaluate metastatic potential, co-immunoprecipitation for protein complex identification, and drug sensitivity testing with chemotherapeutic agents. These polyclonal knockout cells are particularly valuable for investigating colorectal cancer tumor suppression, the impact of circadian rhythm disruption on cancer progression, TGF-??-induced apoptosis, hypoxia response, and preclinical drug efficacy. For further details, please contact Ascent Research.
