The CCNT2 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MES-OV human ovarian carcinoma cell line. These cells carry a targeted disruption of the CCNT2 gene, encoding cyclin T2, the regulatory subunit of P-TEFb. The polyclonal format provides a heterogeneous knockout population for loss-of-function studies, avoiding clonal selection bias. This model is suited for investigating CCNT2-dependent transcriptional regulation in an epithelial cancer context.
MES-OV is a well-characterized human ovarian clear cell carcinoma line established from a patient tumor. It retains epithelial characteristics and tumorigenicity, making it a relevant model for studying ovarian cancer progression, drug resistance, and metastasis. As the host for CCNT2 knockout, MES-OV provides a disease-relevant background to examine P-TEFb-mediated transcriptional elongation in ovarian carcinoma.
Cyclin T2 partners with CDK9 to form the P-TEFb kinase complex, which phosphorylates the C-terminal domain of RNA polymerase II at serine 2, releasing paused polymerase into elongation. P-TEFb also phosphorylates NELF and DSIF, relieving their negative elongation effects. Upstream, P-TEFb is regulated by sequestration in the 7SK snRNP via HEXIM1, chromatin recruitment by BRD4, and NF-??B-driven transcription. In HIV-1 infection, Tat recruits P-TEFb to the TAR element, enhancing viral transcription. Downstream targets include immediate early genes MYC, FOS, and JUN, linking transcriptional elongation to cell growth and stress responses.
In ovarian carcinoma, dysregulated P-TEFb activity drives oncogenic transcriptional programs. The CCNT2 knockout in MES-OV cells enables dissection of cyclin T2’s role in maintaining RNA polymerase II phosphorylation dynamics, NELF/DSIF regulation, and expression of cancer-related genes. This model also facilitates studies of CCNT2 in apoptosis and cell cycle control, pathways often altered in ovarian cancer. Moreover, these cells support HIV-1 Tat transcription research within a human cancer setting.
Typical applications include RNA-seq for global transcription profiling, MTT and annexin V assays for CDK9 inhibitor validation, co-immunoprecipitation of CDK9 to assess P-TEFb integrity, ChIP-qPCR for RNA Pol II Ser2 promoter occupancy, and RT-qPCR for downstream targets MYC and FOS. HIV-1 Tat-dependent transcription can be quantified using reporter assays. This polyclonal knockout model is a versatile tool for CCNT2 functional studies in cancer and virology. For further technical details, contact Ascent Research.
