The KLHL36 Knockout HCT 116 Polyclonal Cells comprise a genetically heterogeneous population of the human colorectal carcinoma line HCT 116 in which the KLHL36 locus has been disrupted using CRISPR/Cas9 technology. This polyclonal knockout pool retains the diversity of independent editing events, avoiding clonal selection biases and providing a robust loss-of-function model. The cells are supplied as a ready-to-use system for interrogating KLHL36 function within the ubiquitin-proteasome pathway in an epithelial colorectal cancer background.
HCT 116 is a well-characterized colon carcinoma cell line with epithelial morphology, widely utilized as a model for colorectal cancer research. Its rapid proliferation and tumorigenic capacity in xenograft assays make it a reproducible platform for examining cellular processes relevant to neoplasia, including proliferation, apoptosis, and drug susceptibility. The line’s genetic background enables studies of oncogenic signaling and tumor suppressor networks in a colonic microenvironment.
KLHL36 encodes a substrate-specific adaptor for the Cullin3-RING E3 ubiquitin ligase (CRL3) complex. It interacts directly with CUL3 and the RING protein RBX1 to recruit E2 ubiquitin-conjugating enzymes, which catalyze the transfer of ubiquitin onto bound substrates. Although the specific targets of KLHL36-mediated ubiquitination are not yet identified, this activity is predicted to regulate the polyubiquitination and subsequent 26S proteasome-dependent degradation of proteins implicated in cell cycle control and stress responses. The KLHL36-CRL3 axis is an integral node within a cascade that includes E1 activating enzymes, E2 conjugating enzymes, and the 26S proteasome, modulating protein homeostasis through precise proteolytic events.
In the context of HCT 116 cells, disruption of KLHL36 offers a focused tool to dissect the contribution of ubiquitin-driven proteolysis to colorectal cancer phenotypes. Because dysregulation of the ubiquitin-proteasome system is a hallmark of many malignancies, this knockout model allows researchers to test how loss of KLHL36 alters degradation of hypothetical substrates that may govern proliferative signaling, checkpoint integrity, or apoptotic sensitivity. The polyclonal nature of the population simulates the cellular heterogeneity observed in tumors, enhancing the translational relevance of experimental outcomes.
This KLHL36 polyclonal knockout pool is compatible with a range of experimental workflows. KLHL36 ablation can be verified by Western blotting and RT-qPCR, while co-immunoprecipitation and ubiquitination assays enable analysis of CRL3 complex assembly and substrate ubiquitination status. Proteasome activity can be monitored using fluorogenic peptide substrates. Functional consequences can be assessed through viability assays (MTT/CCK-8), apoptosis quantification (Annexin V/PI flow cytometry), and cell cycle analysis. Additionally, mass spectrometry-based proteomics may be employed to identify proteins whose stability is altered in the absence of KLHL36. For additional information, please contact Ascent Research.
