The CCNE1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 colorectal carcinoma line, designed to disrupt the CCNE1 gene encoding Cyclin E1. Supplied as a heterogeneous pool, this model allows investigation of Cyclin E1 loss-of-function in a polyclonal context, enabling studies of G1/S transition and cell cycle dysregulation without clonal bias. The product serves as a valuable resource for examining Cyclin E1-associated oncogenic mechanisms and for screening targeted therapies.
The parental HCT 116 cell line is a widely utilized model of human colorectal adenocarcinoma, characterized by a KRAS G13D mutation and microsatellite instability from MLH1 deficiency. These genetic features render HCT 116 cells proficient in MAPK/ERK signaling and defective in DNA mismatch repair, closely mimicking aggressive colorectal cancers. The line??s epithelial morphology and stable karyotype make it suitable for genetic manipulation and subsequent functional assays in cancer biology.
CCNE1 encodes Cyclin E1, the regulatory partner of CDK2. The Cyclin E1/CDK2 complex phosphorylates RB, releasing E2F transcription factors such as E2F1, which drive the G1/S transition and DNA replication. Cyclin E1 expression is activated by E2F1, MYC, ERK/MAPK signaling, and estrogen receptor pathways, and is restrained by CDK inhibitors p21 and p27 and FBXW7-mediated degradation. Downstream, RB hyperphosphorylation enables E2F-mediated transcription of genes including CDC6, and NPAT-dependent histone biosynthesis, promoting S-phase entry and centrosome duplication.
In the HCT 116 background, CCNE1 polyclonal knockout allows dissection of Cyclin E1 function in KRAS-driven, mismatch repair-deficient cells. Cyclin E1 overexpression is frequent in colorectal tumors and linked to chromosomal instability and poor outcomes, so this model assesses transformed cell dependency on Cyclin E1. It supports synthetic lethality studies with CDK inhibitors, revealing vulnerabilities in tumors with constitutive MAPK activation. Such approaches can inform therapeutic strategies targeting cell cycle deregulation in colorectal cancer.
Applications of these polyclonal knockout cells span cancer cell biology, drug discovery, and functional genomics. They are suitable for cell cycle analysis via flow cytometry and BrdU incorporation, examination of downstream signaling by Western blotting for phospho-RB and Cyclin E1 levels, and transcript analysis via RT-qPCR. The model additionally supports colony formation assays and drug sensitivity screens, particularly with CDK inhibitors, to evaluate the role of Cyclin E1 in therapeutic response. For further experimental guidance or product support, please contact Ascent Research.
