The BIRC6 Knockout HCT 116 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal cell population derived from the HCT 116 human colorectal carcinoma cell line, in which the BIRC6 gene has been disrupted. This polyclonal knockout format provides a loss-of-function model that does not rely on single-cell cloning, yielding a heterogeneous population that better reflects genetic variation while maintaining robust target-gene disruption. The polyclonal cells are suitable for studying BIRC6-dependent cellular processes in a cancer-relevant background.
HCT 116 is a widely employed human colorectal carcinoma line harboring an oncogenic KRAS G13D mutation and deficient mismatch repair due to MLH1 deficiency, resulting in microsatellite instability-high (MSI-H) status. These epithelial cells serve as a model for colorectal tumorigenesis and have been extensively used to investigate mechanisms of drug resistance, apoptosis regulation, and tumor cell survival. The genetic background renders HCT 116 cells particularly suited for analyzing DNA damage responses and ubiquitin-proteasome pathway alterations.
BIRC6 encodes an anti-apoptotic E3 ubiquitin ligase that directly ubiquitinates pro-apoptotic factors SMAC (DIABLO) and caspase-9, promoting their proteasomal degradation and suppressing the intrinsic apoptotic cascade. BIRC6 also interacts with HtrA2 and E2 ubiquitin-conjugating enzymes to modulate cell death and survival. Transcriptionally regulated by the oncogene MYC, BIRC6 functions upstream of caspase-3 activation by preventing Apaf-1-mediated cytochrome c-dependent apoptosome formation. In addition to apoptosis, BIRC6 contributes to cytokinesis and DNA repair processes, thus integrating multiple cellular stress responses.
In the HCT 116 context, where BIRC6 expression contributes to apoptosis resistance, the CRISPR/Cas9-mediated knockout of BIRC6 is expected to sensitize cells to apoptotic stimuli, including chemotherapeutic agents. This polyclonal model enables investigation of BIRC6??s role in maintaining viability under DNA-damaging conditions, particularly relevant given the inherent DNA repair defects and MSI-H phenotype. The mixed population reduces clonal bias and allows assessment of BIRC6 loss across a spectrum of genotypes, facilitating robust functional studies in colorectal cancer and other BIRC6-overexpressing malignancies such as breast, prostate, and glioblastoma.
These knockout cells are applicable in a range of experimental settings, including apoptosis mechanism studies using Annexin V staining and caspase activity assays, cell viability and colony formation assays to assess survival, and ubiquitination assays to monitor BIRC6 substrate modification. Western blotting for BIRC6, SMAC, and caspase-9 enables verification of protein-level changes. The model also supports drug resistance research and ubiquitin-proteasome pathway investigation. Researchers can utilize these cells to dissect BIRC6-dependent signaling networks. For additional technical details, please contact Ascent Research.
