The CCNE1 Knockout THP-1 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout population designed to disrupt the CCNE1 gene in THP-1 human monocytic leukemia cells. This product provides a polyclonal loss-of-function model that eliminates cyclin E1 protein without single-cell cloning, ideal for population-based studies of cyclin E1-dependent processes.
THP-1 is a human acute monocytic leukemia cell line derived from the peripheral blood of a one-year-old male. It serves as a widely adopted model for monocyte and macrophage biology, innate immunity, and leukemia research. THP-1 cells can be stimulated with PMA to differentiate into macrophage-like cells, enabling studies in both suspension and adherent formats. Their proliferative capacity and leukemic origin make them particularly suitable for examining the contributions of cell cycle regulators to cancer.
Cyclin E1, encoded by CCNE1, is the regulatory partner of CDK2. Upon activation, cyclin E1?CCDK2 phosphorylates RB1, releasing E2F1 to drive transcription of S-phase genes such as PCNA and MCMs. Cyclin E1 is transcriptionally induced by E2F and MYC, and its levels are controlled by the ubiquitin ligase FBXW7. Negative regulators include CDKN1A (p21) and CDKN1B (p27). The cyclin E1/CDK2 complex integrates signals from growth factor pathways (EGF, estrogen) and is coordinated with the p53 and PI3K/AKT pathways through interactions with CDC25A and SKP2. Dysregulation of cyclin E1 results in unscheduled S-phase entry and genomic instability.
In the THP-1 background, CCNE1 disruption removes the primary G1/S driver, enabling quantification of cyclin E1??s contribution to proliferation and survival. The polyclonal knockout population allows analysis of oncogene addiction, revealing whether THP-1 cells are dependent on cyclin E1 for continued growth. It also permits investigation of compensatory cyclin E2 upregulation and cross-talk with p53-mediated apoptosis. These cells provide a genetically clean system to study how loss of cyclin E1 affects the response to chemotherapeutic agents and targeted therapies.
Researchers can use these knockout cells for in-depth investigation of cyclin E1 function in leukemia. Cell cycle distribution is readily assessed by flow cytometry with propidium iodide staining. Proliferation and viability are quantified using MTS or CellTiter-Glo assays. Cyclin E1 and CDK2 protein levels can be monitored by Western blotting, while mRNA expression is validated by RT-qPCR. The model is also amenable to RNA-seq transcriptome profiling to identify global expression changes upon CCNE1 loss. In drug discovery, the cells support high-throughput screening to identify compounds that exhibit synthetic lethality with cyclin E1 deficiency or that target residual cyclin E2 activity. For additional information, please contact Ascent Research.
