The KBTBD6 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KBTBD6 gene in the HCT 116 colorectal carcinoma line. This heterogeneous pool of edited cells, each with distinct target-site modifications, enables loss-of-function studies without the biases of single-cell cloning, making it ideal for population-level functional assays. The CRISPR/Cas9-mediated gene disruption eliminates KBTBD6 protein expression, establishing a versatile model for investigating its role.
HCT 116 is a well-characterized human epithelial colorectal carcinoma cell line with a near-diploid karyotype. It harbors activating mutations in KRAS (G13D) and PIK3CA, and is deficient in the DNA mismatch repair protein MLH1, resulting in high-frequency microsatellite instability (MSI-H). These genetic features make HCT 116 a widely used model for studying colorectal tumorigenesis, DNA repair deficiencies, and drug responses.
KBTBD6 encodes a substrate adaptor of the Cullin3-RING E3 ubiquitin ligase (CRL3) complex, which includes the scaffold CUL3 and the RING protein RBX1. Within this complex, KBTBD6 specifically recruits p120-catenin (CTNND1) for K48-linked polyubiquitination, tagging it for proteasomal degradation. This process is dependent on CUL3 neddylation by NEDD8 and the activity of E2 enzymes such as UBE2M. By regulating CTNND1 stability, KBTBD6 controls adherens junction integrity and cadherin-mediated cell adhesion, directly impacting epithelial cohesion and motility.
In the HCT 116 colorectal cancer context, KBTBD6 disruption is predicted to stabilize p120-catenin, potentially reinforcing adherens junctions and modulating cell adhesion, migration, and invasion. The co-occurrence of oncogenic KRAS and PIK3CA mutations, along with MSI-H, provides a unique background to investigate how ubiquitin-dependent adhesion regulation intersects with cancer signaling pathways. This polyclonal knockout model thus allows interrogation of KBTBD6??s role in tumor cell dynamics and its potential as a therapeutic vulnerability.
This model supports diverse research applications, including biochemical analysis of CRL3 complex assembly via co-immunoprecipitation and in vitro ubiquitination assays. Protein stability can be assessed by cycloheximide chase, while immunofluorescence microscopy reveals p120-catenin localization at cell junctions. Functional consequences on cell behavior are measured through wound healing, invasion, and adhesion assays. The cells are also amenable to drug library screens targeting the ubiquitin-proteasome system. Standard verification methods include Sanger sequencing, Western blotting, and RT-qPCR. For further information, please contact Ascent Research.
