The KLHDC4 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HEK293T cells, featuring a targeted disruption of the KLHDC4 gene. KLHDC4 encodes the substrate receptor of the CRL2 E3 ubiquitin ligase complex. This polyclonal format provides a heterogeneous pool of knockout cells suitable for functional studies where clonal homogeneity is unnecessary, enabling investigation of KLHDC4-dependent processes in a defined genetic background.
HEK293T is a human embryonic kidney epithelial cell line transformed with SV40 large T-antigen, originating from a female donor. Widely used for recombinant protein expression and viral production due to efficient transfection and robust growth, these cells offer a well-characterized host for studying ubiquitin-mediated degradation pathways.
KLHDC4 serves as a substrate recognition module for the CRL2 complex, interacting with Cullin-2 (CUL2), Elongin B/C (ELOB/ELOC), and RBX1. It specifically binds proteins with C-terminal degron motifs, promoting their ubiquitination and subsequent proteasomal degradation. Upstream regulation involves Cullin-2 neddylation and cellular stress signals, while downstream targets encompass C-terminal degron-containing substrates. This positions KLHDC4 centrally in protein quality control and the ubiquitin-proteasome system, with relevance to cancer and neurodegeneration.
In the HEK293T context, KLHDC4 knockout disrupts CRL2-dependent substrate turnover, providing a tool to dissect degron-mediated proteolysis. The polyclonal nature allows population-level analyses without clonal selection bias, ideal for ubiquitination profiling or proteasome inhibition experiments. This model supports research into how CRL2 activity influences epithelial cell growth and stress responses.
Applications include western blotting, RT-qPCR, ubiquitination assays, cycloheximide chase, proteasome inhibition studies, and immunofluorescence. These cells enable protein degradation analyses, functional genomics, and drug target validation targeting the CRL2 complex. Researchers can combine them with neddylation or proteasome inhibitors to explore ubiquitin-dependent signaling mechanisms. For additional information, contact Ascent Research.
