CCNE2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, engineered for loss-of-function studies of the CCNE2 gene. This polyclonal pool contains cells harboring targeted disruptions at the CCNE2 locus, providing a versatile model for exploring cyclin E2-dependent processes without clonal bias. The use of a polyclonal population allows researchers to assess overall gene function while mitigating effects of clonal variation often seen in single-cell-derived lines.
HAP1 cells are a human near-haploid cell line originally isolated from a patient with chronic myeloid leukemia (CML). Characterized by a fibroblast-like morphology and a near-haploid karyotype, HAP1 cells offer a simplified genetic background that greatly facilitates knockout generation and reduces the complication of redundant gene copies. Their leukemic origin and retention of the BCR-ABL1 fusion make them a relevant platform for studying signaling pathways dysregulated in myeloproliferative disorders and other cancers.
CCNE2 encodes cyclin E2, a regulatory subunit that binds and activates CDK2 to form an active kinase essential for the G1/S transition. The complex phosphorylates RB1, releasing E2F transcription factors such as E2F1 to transcribe S-phase genes including CDC6 and MCM. CCNE2 expression is transcriptionally regulated by E2F and MYC, and restrained by CDK inhibitors p21 and p27 via p53-dependent checkpoints. Cyclin E2 stability is also controlled by the ubiquitin ligase adaptors SKP2 and CKS1B. Disruption of CCNE2 ablates CDK2 kinase activity, preventing RB1 phosphorylation and inducing G1 arrest through sustained E2F inhibition.
The near-haploid nature of HAP1 cells ensures that CRISPR/Cas9-mediated disruption ablates the single functional CCNE2 allele, resulting in a uniform loss of cyclin E2 protein without wild-type compensation. This clean loss-of-function system is especially informative for leukemogenesis studies, as CCNE2 is frequently overexpressed in leukemias and other cancers. Researchers can use this model to probe cyclin E2??s role in DNA replication fidelity, G1 checkpoint integrity, and cellular responses to chemotherapeutic agents or CDK inhibitors.
CCNE2 Knockout HAP1 Polyclonal Cells support a range of cell cycle and cancer research applications. Standard assays include MTS/MTT and colony formation to assess growth dependency; BrdU incorporation and flow cytometry to quantify G1 arrest; and western blotting or RT-qPCR for cyclin E2, phospho-RB1, CDC6, and E2F targets. They are suitable for identifying genetic or chemical modulators of the cell cycle. The polyclonal format is also well-suited for pooled screening and drug response profiling. For further information, please contact Ascent Research.
