KLHL36 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, featuring targeted disruption of the KLHL36 gene. This loss-of-function model impairs KLHL36 protein function, enabling research into ubiquitin-dependent proteostasis and colorectal cancer biology. The polyclonal format provides a heterogeneous genetic background, capturing a range of mutations that better simulate population-level responses compared to isogenic clones.
HT29 cells were established from a primary colorectal adenocarcinoma of a 44-year-old female and serve as a classic model for intestinal epithelial biology. They maintain characteristics such as polarized monolayer formation, mucin secretion, and barrier function, making them a robust platform for studying colorectal cancer pathways, epithelial-to-mesenchymal transition, and oncogene signaling. In the knockout context, these features allow dissection of KLHL36 function in physiologically relevant epithelial processes.
KLHL36 functions as a substrate-specific adaptor within the Cullin3-RING E3 ubiquitin ligase (CRL3) complex. It interacts with Cullin3 and RBX1 to recruit target proteins for ubiquitin conjugation, which directs them to proteasomal degradation. Although the full range of substrates is unknown, KLHL36 is thought to be activated by cellular stress signals and may crosstalk with the Nrf2 antioxidant pathway. By controlling selective protein turnover, KLHL36 influences cell cycle regulation, apoptosis, and redox homeostasis.
Disrupting KLHL36 in HT29 cells permits functional studies of CRL3-dependent degradation in colorectal adenocarcinoma. The knockout may lead to accumulation of substrate proteins, affecting proliferation, migration, and stress responses. Because HT29 cells produce mucin and form barriers, this system is ideal for investigating how ubiquitin-mediated proteolysis impacts epithelial integrity and secretory functions. It also provides a discovery tool for identifying novel KLHL36 substrates via proteomic approaches.
Key applications include knockout validation by western blot and RT-qPCR, proliferation and migration/invasion assays, ubiquitination and proteasome activity measurements, and co-immunoprecipitation with Cullin3 to probe complex assembly. Immunofluorescence enables visualization of substrate accumulation or protein relocalization. These polyclonal knockout cells support both mechanistic studies and drug screening for modulators of the ubiquitin-proteasome system. For further information and ordering, contact Ascent Research.
