The HMBOX1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, engineered for targeted disruption of the HMBOX1 gene. This knockout model eliminates functional HMBOX1 protein expression across a heterogeneous cell pool, enabling bulk-population studies of gene loss without single-cell cloning. The polyclonal format retains genetic diversity while providing a robust loss-of-function system suitable for high-throughput screening, pooled functional genomics, and population-level signaling analyses.
The parental HT29 cell line is a widely used model of human colorectal cancer, originally isolated from a 44-year-old female patient with colorectal adenocarcinoma. These adherent epithelial cells harbor clinically relevant oncogenic mutations, including APC, TP53 (R273H), BRAF V600E, and PIK3CA (E545K), reflecting key driver events in colorectal tumorigenesis. HT29 cells form polarized monolayers and are employed extensively in studies of intestinal epithelial barrier function, cancer cell signaling, and drug response. Their characterized genetic landscape provides a contextually appropriate background for investigating tumor suppressor mechanisms.
HMBOX1 encodes a homeobox transcription factor that acts as a tumor suppressor in colorectal cancer. It directly interacts with RELA and TP53, inhibiting NF-??B activity and promoting p53-dependent apoptosis. HMBOX1 transcriptionally represses CCND1 and MYC, downregulates BCL2, and induces CDKN1A and BAX. Additionally, it suppresses Wnt/??-catenin signaling by disrupting CTNNB1/TCF4 complexes, reducing EMT-related gene expression. Upstream regulators include NF-??B, p53, and microRNAs miR-9, miR-19a, and miR-124.
In HT29 cells, which carry activating mutations in BRAF and PIK3CA and defective APC and TP53, HMBOX1 knockout exacerbates the transformed phenotype, making this model valuable for dissecting compensatory tumor-suppressive mechanisms. Loss of HMBOX1 relieves repression on NF-??B and Wnt pathways, enhancing proliferation, survival, and migratory capacity. This polyclonal knockout population allows researchers to study HMBOX1??s role in maintaining epithelial integrity and suppressing EMT in a colorectal cancer background. It also provides a platform to evaluate how HMBOX1 deficiency cooperates with existing oncogenic lesions to drive malignancy.
These cells are ideal for a broad range of functional assays, including western blot, RT-qPCR, RNA-seq, and ChIP-qPCR. NF-??B and Wnt/??-catenin reporter assays directly measure pathway activation. Functional readouts such as colony formation, wound healing, and apoptosis assays are well suited. Co-immunoprecipitation confirms disrupted HMBOX1 interactions. The polyclonal population facilitates drug screening and pooled CRISPR studies. For detailed technical specifications, please contact Ascent Research.
