The ANAPC7 Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HT29 cells carrying targeted disruption of the ANAPC7 gene. This pool of edited cells provides a heterogeneous loss-of-function model for the anaphase-promoting complex/cyclosome (APC/C) core subunit ANAPC7, enabling robust interrogation of APC/C-dependent mitotic regulation. As a polyclonal knockout population, it reflects the average effect of diverse editing events across the culture, making it suitable for bulk functional genomic studies and high-throughput screens in a colorectal adenocarcinoma background.
The parental HT29 cell line is a well-characterized human colon adenocarcinoma model derived from a female patient. These epithelial cells harbor mutations in APC and TP53, are proficient in mismatch repair, and retain the capacity for enterocytic differentiation, making them a standard tool for intestinal epithelial biology and colorectal cancer research. Their genetic profile recapitulates key features of chromosomal instability (CIN) and Wnt pathway dysregulation observed in many colorectal tumors, providing a physiologically relevant platform for studying tumor cell cycle control.
ANAPC7 encodes the APC7 subunit of the APC/C E3 ubiquitin ligase, which governs mitotic progression by targeting cell cycle regulators for proteasomal degradation. Its activity is controlled by the co-activators CDC20 and CDH1, mitotic kinases CDK1/Cyclin B and PLK1, and the spindle assembly checkpoint proteins MAD2 and BUBR1. Key substrates of the ANAPC7-containing APC/C include securin, cyclin B1, Aurora A, PLK1, and CDC20 itself. Together with the E2 enzymes UBE2C and UBE2S, the complex ensures orderly mitotic exit. Disruption of ANAPC7 abrogates APC/C ligase function, leading to stabilization of these regulators, mitotic arrest, and chromosomal instability.
In the HT29 background, ANAPC7 knockout exacerbates the inherent genomic instability driven by APC and TP53 mutations, generating a powerful cell model to dissect the mechanisms linking APC/C dysfunction to colorectal tumorigenesis. By eliminating the major mitotic regulator ANAPC7, this model mimics defects in the ubiquitin-mediated proteolysis pathway and allows for dissection of APC/C-dependent versus independent cell cycle checkpoints. The interplay between ANAPC7 loss and the spheroid-forming, differentiation-competent nature of HT29 cells further enables studies of how mitotic errors influence tumor cell plasticity and drug response in a CIN-positive context.
This polyclonal knockout cell population is ideally suited for investigations into APC/C biology, cell cycle control, and colorectal cancer pathophysiology. Researchers can employ it in flow cytometry for cell cycle distribution analysis, Western blotting and immunofluorescence to assess mitotic protein stability and subcellular localization, apoptosis assays to evaluate mitotic catastrophe, colony formation and drug sensitivity assays for functional studies, and RT-qPCR to probe transcriptional changes. It serves as a valuable platform for screening mitotic inhibitors or combining with standard chemotherapeutics. For further details on product specifications, validation, and pricing, please contact Ascent Research.
