The KLHL26 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the KLHL26 gene in the human HAP1 cell line. This product enables loss-of-function studies of KLHL26, a substrate adaptor protein for the CUL3-based E3 ubiquitin ligase complex. By targeting the KLHL26 locus, researchers can investigate its role in ubiquitin-mediated proteolysis and protein quality control. The polyclonal nature of the knockout population avoids clonal artifacts and preserves genetic heterogeneity, making it a versatile tool for functional genomics and pathway analysis. The cells are provided as a ready-to-use mixed population, suitable for immediate expansion and downstream applications.
The host cell line, HAP1, is a near-haploid, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its adherent growth and stable, simplified karyotype make it an ideal model for genetic perturbation studies, as the reduced gene copy number facilitates unambiguous phenotypic analysis of loss-of-function mutations. HAP1 cells retain essential signaling pathways and cellular machinery, including a functional ubiquitin-proteasome system, enabling physiologically relevant investigations. The cell line’s leukemic origin also renders it valuable for studying cancer-associated processes, particularly those involving protein homeostasis and oncogenic signaling.
KLHL26 functions as a substrate-specific adaptor for the CUL3-RBX1 E3 ubiquitin ligase complex, recruiting target proteins for polyubiquitination and subsequent proteasomal degradation. Through its BTB domain, KLHL26 homodimerizes and interacts with CUL3, while RBX1 facilitates ubiquitin transfer to substrates. Although the precise downstream ubiquitination targets of KLHL26 remain unknown, its activity is regulated by CUL3 neddylation, a post-translational modification that activates the ligase complex. By mediating substrate recognition, KLHL26 plays a critical role in maintaining protein homeostasis and regulating cellular processes linked to turnover of key signaling factors, potentially influencing pathways relevant to cancer cell proliferation and survival.
In the HAP1 context, disruption of KLHL26 provides a powerful system for dissecting the ubiquitin-proteasome network in a near-haploid genetic background. This knockout model is particularly advantageous for studying the functional consequences of impaired substrate adaptor activity, as HAP1 cells allow rapid generation of homozygous-like gene disruptions without extensive clone isolation. Combined with the cell line’s leukemic heritage, the model offers a platform to investigate how KLHL26-mediated proteolysis may contribute to tumorigenesis, protein quality control defects, and cellular responses to proteotoxic stress. The polyclonal population also supports pooled screening approaches to identify synthetic lethal interactions or drug sensitivities associated with KLHL26 loss.
Typical research applications include biochemical analysis of ubiquitination events via Western blotting and ubiquitination assays, identification of KLHL26 interaction partners through co-immunoprecipitation, and functional proteasome activity measurements. Cell-based assays such as proliferation and viability screens can be coupled with RNA-seq to map transcriptional changes upon KLHL26 ablation. These applications are instrumental for uncovering KLHL26 substrates, validating drug targets within the ubiquitin-proteasome system, and exploring the protein homeostasis landscape in leukemia and other cancers. For further details, pricing, or technical support, please contact Ascent Research.
