The CCNE1 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CCNE1 gene in Raji B lymphocytes. This loss-of-function model is produced through CRISPR/Cas9-mediated gene disruption, yielding a genetically diverse pool of edited cells without clonal selection. By eliminating functional Cyclin E1 expression, the model provides a powerful system for investigating Cyclin E1-dependent processes in cell cycle regulation and oncogenic signaling.
The host Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived B lymphocyte line. It serves as a well-established model for humoral immunity and antigen presentation, with constitutive activation of survival and proliferation pathways driven by EBV. This background makes Raji cells particularly relevant for studying the molecular mechanisms of lymphomagenesis and viral oncogenesis, offering a disease-relevant context for CCNE1 knockout studies.
The CCNE1 gene encodes Cyclin E1, the regulatory subunit of CDK2 essential for G1/S transition and DNA replication. Its expression is driven by E2F and MYC, and activated by MAPK and PI3K/AKT signaling. Cyclin E1-CDK2 complexes phosphorylate Rb, releasing E2F to induce S-phase genes like CDC6 and CDT1. Negative regulation occurs via CDK inhibitors p21/p27 and Fbxw7-mediated degradation. Thus, CCNE1 integrates signals from pRb, p53, and mTOR pathways.
In Raji cells, CCNE1 knockout disrupts Cyclin E1-CDK2 complex formation, attenuating Rb phosphorylation and E2F-driven transcription, potentially causing G1 arrest or apoptosis. As CCNE1 is frequently overexpressed in ovarian, breast, and gastric cancers and linked to poor Burkitt lymphoma prognosis, this model enables dissection of Cyclin E1 oncogenic mechanisms in B lymphocytes. It also permits study of cross-talk with p53 and pRb tumor suppressor pathways.
Applications include cell cycle analysis by flow cytometry, proliferation and apoptosis assays, and Western blotting for phospho-Rb and CDK2. The model supports drug sensitivity testing, colony formation assays, and RT-qPCR for E2F targets. Additionally, the model facilitates studies of compensatory CDK4/6-Cyclin D or p16 pathway alterations. Lymphoma-specific studies can probe EBV-kinase interactions, and functional genomics screens utilize the polyclonal population. For further information, contact Ascent Research.
