The CCNE1 Knockout HT29 Polyclonal Cells represent a rigorously validated CRISPR/Cas9-edited polyclonal knockout cell population for the CCNE1 gene, derived from the HT29 human colorectal adenocarcinoma cell line. This product consists of a heterogeneous pool of gene-disrupted cells, avoiding clonal selection to better recapitulate the genetic diversity observed in tumor populations. Supplied as a ready-to-use cell suspension, it facilitates a wide array of downstream applications in oncology research, particularly for investigating cell cycle regulation and targeted therapy response.
The HT29 cell line is a widely used model of human colon adenocarcinoma, originally isolated from a primary colorectal tumor. These epithelial cells carry mutations in the APC and TP53 tumor suppressor genes, recapitulating common genetic alterations in sporadic colorectal cancer. HT29 cells exhibit robust proliferation and retain a differentiated intestinal phenotype, providing a physiologically relevant context for studying oncogenic signaling and therapeutic responses. The mutant TP53 background abrogates DNA damage checkpoints, accentuating dependence on G1/S regulatory machinery.
Cyclin E1 (CCNE1) encodes the regulatory subunit of CDK2, driving the G1/S transition. Its expression is controlled by transcription factors E2F1 and MYC, with input from RAS and EGFR pathways. The CCNE1-CDK2 complex phosphorylates RB1, liberating E2F transcription factors to execute DNA replication programs. Activity is modulated by CDK inhibitors CDKN1A (p21) and CDKN1B (p27). Overexpression of CCNE1, commonly observed in cancer, leads to accelerated cell cycle progression and genomic instability, underpinning its oncogenic role.
In the HT29 background, with mutant APC and TP53, CCNE1 knockout enables dissection of its contribution to uncontrolled proliferation and tumor maintenance. This model is particularly suited for studying dysregulated pathways in colorectal cancer, including p53, PI3K-Akt, and DNA replication. The polyclonal format allows assessment of CCNE1-dependent vulnerabilities across a heterogeneous population, facilitating identification of therapeutic targets relevant to colorectal, breast, ovarian, and gastric cancers.
Polyclonal CCNE1 knockout HT29 cells enable cell cycle profiling via flow cytometry, western blot analysis of Cyclin E1/CDK2/RB1 phosphorylation, and proliferation assays such as MTT or BrdU incorporation. Researchers can perform RT-qPCR to measure CCNE1 and downstream E2F target transcripts. Colony formation and apoptosis assays further assess proliferative potential and cell death pathways. These tools support validation of putative therapeutic targets, mechanistic investigation of CDK inhibitors, and discovery of synthetic lethal interactions in colorectal cancer research. For further technical assistance, please contact Ascent Research.
