The CCNE1 Knockout A-549 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma cell line, featuring disruption of the CCNE1 gene through targeted gene editing. This polyclonal population provides a heterogeneous loss-of-function model for studying Cyclin E1 functions, suitable for functional genomics and drug discovery applications without the constraints of single-cell clones.
The A-549 host cell line is an epithelial cell model of non-small cell lung cancer (NSCLC), originally established from a 58-year-old Caucasian male with lung adenocarcinoma. These cells are widely used to investigate lung cancer biology, drug response, and signal transduction, making them a clinically relevant platform for CCNE1 knockout studies.
Cyclin E1, encoded by CCNE1, activates CDK2 to promote G1/S transition, phosphorylating RB1 and releasing E2F transcription factors, which drive expression of DNA replication genes like CDC6 and MCM proteins, and histone biosynthesis. CCNE1 is transcriptionally induced by E2F and MYC, and further regulated by RAS/MAPK and PI3K/AKT growth signals. Its activity is restrained by CDK inhibitors p21 and p27, and protein stability is controlled by FBXW7-mediated ubiquitination. In A-549 cells, CCNE1 knockout disrupts CDK2 activation, leading to RB hypophosphorylation, E2F sequestration, and cell cycle arrest, thus blocking proliferation.
In lung adenocarcinoma, CCNE1 amplification or overexpression is linked to aggressive disease. Knocking out CCNE1 in A-549 cells impairs G1/S progression, reduces proliferation, and may induce apoptosis under oncogenic stress. This polyclonal knockout model enables dissection of cell cycle dysregulation in NSCLC and evaluation of CDK-targeted therapies, as well as studies on compensatory mechanisms that may emerge upon Cyclin E1 loss.
Key applications include cell cycle analysis, drug screening, CDK inhibitor testing, and functional studies in lung adenocarcinoma. Assays commonly employed are western blotting for CCNE1, CDK2, RB1, pRB, and p21; RT-qPCR; flow cytometry for cell cycle distribution; proliferation assays (MTT, BrdU); apoptosis assays (Annexin V); colony formation; and xenograft tumor growth. For further details, please contact Ascent Research.
