The HTATIP2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HTATIP2 gene in the HT29 human colorectal adenocarcinoma cell line. This gene-disrupted pool enables robust loss-of-function studies of the tumor suppressor HTATIP2, which promotes apoptosis, inhibits angiogenesis, and regulates metabolism. The polyclonal format preserves heterogeneous editing outcomes, avoiding clonal selection artifacts and enhancing reproducibility.
HT29 cells, originally isolated from a primary colon adenocarcinoma of a 44-year-old Caucasian female, are a well-established model of human intestinal epithelium and colorectal carcinoma. These cells harbor clinically relevant mutations, including BRAF(V600E), mutant p53, and defective APC, which drive aberrant signaling networks. Their adherent epithelial morphology and tumorigenic properties make them invaluable for investigating colon cancer biology and therapeutic responses.
HTATIP2 functions as a tumor suppressor downstream of p53 and E2F1, with its expression induced by DNA damage and oxidative stress. In the intrinsic apoptotic pathway, HTATIP2 promotes Bax oligomerization, leading to mitochondrial outer membrane permeabilization, cytochrome c release, and caspase-9 activation. Beyond apoptosis, HTATIP2 serves as a transcriptional corepressor, directly binding to TBP and NOL8 to repress transcription of the pro-angiogenic factor VEGF. Furthermore, it influences glucose metabolism by modulating the expression of glucose transporters, thereby impacting cellular energy homeostasis.
Disruption of HTATIP2 in HT29 cells abrogates its tumor-suppressive functions, potentially enhancing cell survival, proliferation, angiogenic potential, and metabolic reprogramming. This knockout model is particularly valuable for dissecting HTATIP2’s role in colon cancer progression in the context of p53 and BRAF mutations, including p53-independent apoptotic regulation and metastatic signaling. The loss of HTATIP2 may mirror key hallmarks of aggressive colorectal tumors, making these cells a powerful tool for mechanistic studies.
These polyclonal knockout cells support diverse experimental applications, such as apoptosis assays using annexin V and caspase-9 activation readouts, cell viability and proliferation studies, and migration/invasion assays. Co-immunoprecipitation enables interaction studies with TBP and NOL8, while RT-qPCR quantifies downstream targets like Bax and VEGF. Metabolic profiling via glucose uptake assays further elucidates HTATIP2’s role in cancer metabolism. For in vivo studies, these cells are suitable for tumor xenograft models to assess metastatic potential and drug sensitivity. For further details, contact Ascent Research.
