The KLHL36 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human near-haploid cell line, intended for advanced ubiquitin-proteasome research and functional genomics. This product consists of a heterogeneous pool of cells harboring disruptions at the KLHL36 locus, generated through CRISPR/Cas9-mediated gene targeting, leading to loss of KLHL36 protein function across the population. By avoiding single-cell cloning, this polyclonal format preserves genetic diversity and offers a robust model for studying KLHL36-dependent processes in a pooled context, reflecting broader cellular responses.
The HAP1 host cell line is a chronic myeloid leukemia (CML)-derived near-haploid cell line, originally isolated from the KBM-7 line. It carries the BCR-ABL1 oncogene and displays an adherent morphology. Crucially, its near-haploid karyotype, with only one copy of most genes, facilitates efficient CRISPR/Cas9-mediated knockout and makes it an ideal platform for haploid genetic screens. The CML origin provides a disease-relevant background for investigating kinase-driven signaling and protein homeostasis, particularly in the context of leukemia biology.
KLHL36 functions as a substrate adaptor for Cullin3-RING E3 ubiquitin ligase complexes, serving as a substrate recognition component. Within these complexes, Cullin3 acts as a scaffold, interacting with the RING finger protein RBX1 and E2 ubiquitin-conjugating enzymes to catalyze ubiquitin transfer onto specific substrates recruited by KLHL36. This process targets proteins for proteasomal degradation, thereby regulating their cellular abundance. The core pathway includes CUL3, RBX1, E2 enzymes, and the 26S proteasome. Although the upstream regulators and specific downstream targets of KLHL36 remain unidentified, its role places it within the ubiquitin-proteasome system, influencing protein quality control and cell cycle regulation through post-translational protein turnover.
In the HAP1 BCR-ABL1-positive CML setting, disruption of KLHL36 enables dissection of Cullin3-dependent ubiquitination pathways in a leukemia-relevant context. The ubiquitin-proteasome system is essential for proteostasis in cancer cells, and aberrant E3 ligase activity can contribute to tumorigenesis. This polyclonal knockout model permits investigation of how KLHL36 loss affects global protein ubiquitination, cell cycle progression, and stress responses in a haploid background. It is particularly valuable for haploid genetic screens aimed at uncovering synthetic lethal interactions or therapeutic vulnerabilities associated with BCR-ABL1-driven malignancies.
This KLHL36 knockout product supports diverse applications, including functional genomics, ubiquitination studies, and protein degradation research. Researchers can employ it in CRISPR essentiality screens to assess genetic dependencies, co-immunoprecipitation experiments to probe interactions with CUL3 or RBX1, and western blotting to monitor candidate substrate levels. Ubiquitination assays detect changes in ubiquitin conjugation patterns, while cell viability assays evaluate the impact on proliferation and drug response. The polyclonal format is optimal for pooled screening strategies and broad mechanistic studies. For additional technical details, please contact Ascent Research.
