The C15orf40 Knouckout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 colorectal adenocarcinoma line. This loss-of-function model is designed for the functional investigation of the uncharacterized C15orf40 gene. The polyclonal format, produced without single-cell cloning, yields a heterogeneous mixture of knockout cells, which is advantageous for pooled screening approaches and bulk phenotypic assays, as it provides inherent biological replicates and robust statistical power. CRISPR/Cas9-mediated disruption of the C15orf40 locus impairs protein expression, enabling researchers to dissect the gene??s role in cancer-relevant processes.
HT29 cells were established from a colon adenocarcinoma of a 44-year-old Caucasian female and are characterized by well-defined oncogenic mutations, including APC truncations, TP53 loss-of-function, BRAF V600E, and PIK3CA activating mutations. These alterations result in constitutive activation of the Wnt/??-catenin, MAPK, and PI3K/AKT signaling pathways, which collectively drive proliferation, survival, and transformation. HT29 cells maintain an epithelial phenotype, retain the ability to undergo intestinal differentiation and mucus production, and are widely employed as a model system for studying colorectal carcinoma biology and drug responses.
The C15orf40 gene encodes a protein containing a DUF4557 domain of unknown function, with no validated enzymatic, signaling, or protein interaction motifs reported. To date, no upstream regulatory factors, downstream targets, or interacting partners have been experimentally defined, and the gene is not assigned to any canonical biological pathway. Although transcript expression changes have been documented in colorectal and hepatocellular carcinomas, the molecular basis of its involvement remains elusive. This CRISPR knockout model thus constitutes a valuable platform for the initial functional characterization and deorphanization of C15orf40.
Deploying C15orf40 knockout within the HT29 genetic background permits functional interrogation in a context of concurrent pathway dysregulation. The established activation of Wnt, MAPK, and PI3K/AKT pathways in these cells creates a sensitized environment where the impact of C15orf40 loss on proliferation, apoptosis, migration, and chemosensitivity can be systematically assessed. Comparative phenotypic analyses between the knockout pool and parental HT29 cells can illuminate whether C15orf40 modulates these oncogenic networks or contributes to the transformed phenotype through independent mechanisms.
Typical research applications span functional genomics, cancer cell biology, and drug sensitivity profiling. Standard assays such as EdU proliferation, MTT viability, colony formation, Transwell migration/invasion, and Annexin V/PI apoptosis detection are directly applicable. Western blotting for EMT markers (E-cadherin, vimentin), transcriptome analysis by RNA-seq, and drug sensitivity testing with 5-fluorouracil and oxaliplatin enable detailed mechanistic studies. This model thereby facilitates the identification of C15orf40-dependent vulnerabilities in colorectal cancer. For technical inquiries or custom applications, please contact Ascent Research.
