The CCDC90B Knockout HT29 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the CCDC90B gene in the HT29 human colorectal adenocarcinoma cell line. This polyclonal pool contains a heterogeneous mixture of cells with distinct editing events at the target locus, providing a robust loss-of-function model without clonal selection. The knockout model enables systematic investigation of CCDC90B-dependent mitochondrial functions in an epithelial cancer background. Gene disruption was achieved using CRISPR/Cas9 technology, and the resulting polyclonal population retains the parental line’s fundamental characteristics while ablating CCDC90B expression. This reagent is suited for assays requiring mixed genotypes, such as pooled functional screens or studies assessing population-level phenotypes.
HT29 cells are a well-established human colorectal adenocarcinoma line with epithelial morphology, widely employed as a model for colorectal cancer research. These cells recapitulate key features of intestinal epithelial biology, including polarized architecture, mucin production, and active oncogenic signaling pathways such as Wnt/??-catenin. The HT29 background provides a clinically relevant context for examining mitochondrial contributions to tumor metabolism, proliferation, and therapy resistance. As an adherent line with robust growth characteristics, HT29 supports a broad range of downstream applications, from high-resolution imaging to metabolic flux analysis. Its origin from a primary colonic tumor ensures that findings in this system retain translational relevance for colorectal adenocarcinoma biology.
CCDC90B encodes a mitochondrial inner membrane protein that plays a critical role in cristae organization and respiratory chain complex III assembly. Mechanistically, CCDC90B interacts with components of the MICOS complex (including UQCC3) and the dynamin-related GTPase OPA1 to maintain cristae junction architecture. Its activity is regulated by upstream signals such as HIF1A and nutrient deprivation, linking cristae remodeling to cellular metabolic status. Loss of CCDC90B disrupts these interactions, leading to cristae disorganization, diminished mitochondrial membrane potential, and impaired oxidative phosphorylation. Consequently, CCDC90B knockout sensitizes cells to intrinsic apoptosis through enhanced cytochrome c release and downstream caspase activation, with BAX and BAK serving as key executioners. The protein also functionally interfaces with mitochondrial fusion factors MFN1 and MFN2, integrating cristae dynamics with the mitochondrial network.
The abrogation of CCDC90B in HT29 cells creates a powerful model to dissect mitochondrial dysfunction in colorectal cancer. Colorectal tumors often exhibit metabolic reprogramming driven by HIF1A and nutrient availability, making the CCDC90B-dependent cristae pathway a node of potential vulnerability. In this polyclonal knockout population, the combined effects of disrupted oxidative phosphorylation and heightened apoptotic sensitivity can be interrogated under physiologically relevant stresses such as hypoxia or glucose limitation. The epithelial nature of HT29 further permits studies on how mitochondrial structure?Cfunction relationships impact tumor cell differentiation, invasion, or response to chemotherapeutic agents. This model thus bridges molecular mitochondrial biology and colorectal cancer pathophysiology.
Researchers can employ these CCDC90B knockout HT29 polyclonal cells in a variety of advanced experimental workflows. Assays to confirm target disruption include quantitative Western blotting for CCDC90B, while functional characterization can involve mitochondrial membrane potential measurements using JC-1 or TMRE dyes. Apoptotic priming is readily assessed by Annexin V staining and caspase activation assays, and metabolic consequences are quantifiable via oxygen consumption rate (OCR) analysis. Cristae morphology can be visualized by immunofluorescence staining for OPA1 and DRP1. Additionally, long-term effects on cell growth and clonogenic survival are measurable using colony formation assays. These applications make the product ideal for studies on mitochondrial contributions to drug sensitivity, resistance, and apoptosis signaling in colorectal cancer. For further details or technical support, please contact Ascent Research.
