The HLTF Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population in which the HLTF (helicase-like transcription factor) gene has been disrupted within the HT29 human colorectal adenocarcinoma background. This polyclonal pool offers a genetically heterogeneous loss-of-function model, enabling robust investigation of HLTF-dependent biological processes without the selection bias of clonal isolates.
The parental HT29 cell line is a widely used epithelial model derived from a colorectal adenocarcinoma, retaining enterocytic differentiation potential and serving as a standard platform for intestinal barrier function, colorectal cancer biology, and drug response studies. The line??s well-characterized growth properties and signaling make it particularly suitable for evaluating tumor suppressor gene functions.
HLTF functions as a DNA helicase and chromatin remodeler critical for DNA damage tolerance and genome maintenance. It is recruited to stalled replication forks by RAD18-ubiquitinated PCNA following ATR activation, where it promotes fork reversal and template switching to enable error-free lesion bypass. In this process, HLTF interacts with PCNA, RAD18, RAD51, and BRCA2, facilitating RAD51 loading and replication fork stability. As a member of the RAD6-RAD18 pathway, HLTF opposes translesion synthesis, and its levels are controlled by APC/C-mediated proteolysis. Its tumor suppressor role is exemplified by frequent promoter hypermethylation-associated silencing in colorectal and other cancers.
In the HT29 colorectal carcinoma context, HLTF loss is particularly relevant given its epigenetic silencing in colorectal tumors. This polyclonal knockout model recapitulates HLTF deficiency, enabling studies of how its absence affects DNA repair, replication stress responses, and chemosensitivity. The HT29 background provides a malignant epithelial environment in which to dissect HLTF??s impact on cell proliferation, apoptosis, and differentiation, offering insights into its tumor-suppressive mechanisms and the consequences of its inactivation in intestinal cancers.
Researchers can utilize this model for DNA damage assessments (??-H2AX foci, Comet assay), replication stress analysis (DNA fiber labeling), cell viability and clonogenic survival assays with cisplatin or 5-FU, cell cycle analysis, and apoptosis studies. Gene disruption can be confirmed by RT-qPCR and western blotting. This polyclonal knockout cell population supports chemosensitivity screening, tumor suppressor research, and pathway dissection. For inquiries or ordering, please contact Ascent Research.
