The CCNB2 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population targeting CCNB2 in HCT 116 colorectal carcinoma cells. This pooled knockout model is generated by transient Cas9?CgRNA delivery, producing a heterogeneous population of loss-of-function alleles. The polyclonal format minimizes clonal selection bias and is suited for high-throughput phenotypic screening, enabling robust investigation of CCNB2-dependent pathways in a disease-relevant context for functional genomics and drug target validation.
The HCT 116 cell line is a male human colorectal adenocarcinoma model with epithelial morphology, harboring a KRAS G13D mutation and MLH1-deficient MSI-H status. These features mirror a subset of colorectal cancers with defective DNA mismatch repair, making it an appropriate host for studying genotype-specific mitotic vulnerabilities. The line is extensively used in drug sensitivity profiling, xenograft models, and molecular oncology research.
CCNB2 encodes cyclin B2, a CDK1 regulatory subunit that drives G2/M progression. Cyclin B2?CCDK1 complexes phosphorylate substrates for mitotic spindle assembly and nuclear envelope breakdown. Transcription is activated by FOXM1, E2F1, and NF-Y, and repressed by p53 at the G2/M checkpoint. Downstream targets include PLK1, CDC25C, and the inhibitory kinases WEE1/MYT1, forming feedback loops that regulate mitotic timing. Cyclin B2 also interacts with microtubule-associated proteins to localize activity to the spindle. Disruption of CCNB2 in HCT 116 leads to mitotic arrest and loss of mitotic fidelity.
In the HCT 116 MSI-H background, CCNB2 knockout allows dissection of mitotic signaling in colorectal cancer. MSI-H tumors often show mitotic regulator dysregulation and sensitivity to spindle poisons, providing a system for synthetic lethality studies. The KRAS G13D mutation further modulates proliferation and survival pathways, enhancing the model??s relevance for combination therapy screening. The polyclonal architecture ensures that observed phenotypes are consistent across diverse editing outcomes, increasing data reproducibility in pooled assays.
Applications include cell cycle analysis via flow cytometry, MTT and colony formation assays, and immunofluorescence for phospho-histone H3. Western blotting for cyclin B2 and phospho-CDK1 confirms target-gene disruption. Time-lapse microscopy captures mitotic progression defects. These tools support mitotic inhibitor screening and evaluation of G2/M-targeted agents. The cells also enable CRISPR-based phenotypic and genetic interaction studies. For further information, please contact Ascent Research.
