AIF1L Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the HT29 human colorectal adenocarcinoma cell line, engineered for loss-of-function studies of the AIF1L gene. This knockout model provides a genetically heterogeneous pool of edited cells, enabling pooled functional analyses of AIF1L-dependent processes without clonal selection.
The HT29 cell line is a well-established adherent epithelial model originating from a human colorectal adenocarcinoma, characterized by mutations in the tumor suppressors APC and TP53, which simulate key oncogenic features. These cells exhibit an epithelial morphology and are widely employed as a reproducible intestinal epithelial platform for colorectal cancer research, drug screening, and mechanistic studies of gut inflammation.
The AIF1L gene encodes an actin-bundling protein that orchestrates cytoskeletal dynamics, cell migration, and inflammatory response modulation. Mechanistically, AIF1L is activated by upstream signals including TNF-alpha, IL-1beta, LPS, and NF-kB, and it operates within the Cdc42/Rac1 signaling cascade that converges on actin remodeling. It directly interacts with actin and calcium ions and regulates downstream effectors such as actin polymerization, focal adhesion turnover, and cofilin-mediated filament severing. Through these interactions, AIF1L influences cell motility and the production of inflammatory cytokines.
In the HT29 colorectal cancer context, disruption of AIF1L is predicted to impair actin cytoskeleton integrity, thereby diminishing cellular migration and invasive capacity??hallmarks of tumor progression. The inherent APC and TP53 mutations in HT29 cells create a tumorigenic background that enhances the relevance of this knockout for probing how actin-bundling alterations interact with oncogenic signaling. Additionally, loss of AIF1L may perturb NF-kB-mediated cytokine expression, offering a model to explore the cross-talk between cytoskeletal reorganization and inflammatory signaling in colorectal cancer and inflammatory bowel disease.
This polyclonal knockout cell population is suitable for a range of assays: analyzing AIF1L??s role in colorectal cancer cell motility using wound healing and transwell invasion assays; investigating inflammatory pathways via western blotting, RT-qPCR, and flow cytometry for cytokines; and visualizing actin cytoskeleton changes through immunofluorescence. The model also supports drug sensitivity studies targeting cytoskeletal regulators and can be applied to research on transplant rejection and autoimmune disorders where AIF1L-mediated inflammation is implicated. For further technical information, please contact Ascent Research.
