The CCNT2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CCNT2 gene in human HT29 colorectal adenocarcinoma cells. This product offers a heterogeneous cell pool with disrupted CCNT2 alleles, enabling loss-of-function analyses without clonal isolation. The polyclonal format preserves genetic diversity, which is advantageous for transcriptional profiling and drug-response screens in a cancer-relevant context.
HT29 cells, derived from a 44-year-old Caucasian female with colorectal adenocarcinoma, are a well-established model for colorectal cancer and intestinal epithelial biology. These adherent epithelial cells can undergo differentiation under specific conditions, making them suitable for studies of tumor cell signaling and drug metabolism. The p53-mutant background further enhances their relevance in oncogenic pathway research, and when coupled with CCNT2 disruption, they provide a robust system for investigating P-TEFb-mediated transcriptional regulation in colorectal cancer.
CCNT2 encodes Cyclin T2, the regulatory subunit of the P-TEFb complex that controls RNA polymerase II (Pol II) pause release. Cyclin T2 binds CDK9, which then phosphorylates the Pol II CTD at serine 2 and negative elongation factors DSIF and NELF, driving productive elongation. P-TEFb activity is regulated by the 7SK snRNP (LARP7, MEPCE) and HEXIM1, which inhibit kinase function, while HIV-1 Tat and BRD4 competitively activate P-TEFb. Key transcriptional targets include MYC and FOS, linking Cyclin T2 to cell cycle progression.
In HT29 cells, CCNT2 disruption is valuable for studying how transcriptional dysregulation contributes to colorectal tumorigenesis. P-TEFb is often exploited by oncogenic pathways to sustain high expression of proliferative genes. Ablating Cyclin T2 impairs CDK9-dependent Pol II phosphorylation, potentially reducing MYC-driven transcription and affecting cell proliferation and survival, thereby enabling dissection of P-TEFb-dependent oncogenic mechanisms and evaluation of CDK9 inhibitors like flavopiridol.
Research applications include transcriptional elongation studies, HIV transcription research, and colorectal cancer profiling. Typical assays are western blotting for Cyclin T2 and CDK9, ChIP-qPCR for Pol II occupancy, RNA-seq, RT-qPCR, CDK9 kinase assays, co-immunoprecipitation of P-TEFb components, HIV-1 LTR reporter assays, and cell viability assays. This model supports drug screening and mechanistic investigation of CDK9-dependent gene regulation. For inquiries, contact Ascent Research.
