The KLHL18 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function analysis of the KLHL18 gene in a near-haploid human cell background. This product is generated via CRISPR/Cas9-mediated gene disruption, resulting in a mixed population of edited cells with targeted disruption of KLHL18, providing a robust model to study the gene??s role in ubiquitin-dependent proteasomal degradation. The polyclonal format eliminates the need for single-cell cloning, preserving genetic diversity and facilitating large-scale functional studies.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia cell line and maintain a near-haploid karyotype, which simplifies genetic screens and loss-of-function analyses. These adherent cells exhibit a fibroblast-like morphology and originate from a male donor. Importantly, HAP1 cells express the BCR-ABL fusion protein, characteristic of chronic myeloid leukemia, making them particularly relevant for cancer research and functional genomics. Their robustness and ease of culture support a wide range of in vitro assays, including high-throughput screening and detailed mechanistic investigations.
KLHL18 functions as a substrate adaptor within the Cullin3-RING E3 ubiquitin ligase complex (CRL3), where it interacts directly with Cullin3 and RBX1 to recruit specific substrates for ubiquitination. This polyubiquitination marks target proteins for recognition and degradation by the 26S proteasome. Although the full repertoire of KLHL18 substrates remains largely undefined, potential targets are thought to include proteins involved in cytoskeletal dynamics and cell cycle regulation. Post-translational modification via ubiquitin chains is a critical regulatory mechanism, and KLHL18-mediated ubiquitination contributes to the control of protein turnover, impacting diverse cellular processes. Upstream regulation of KLHL18 expression is not extensively characterized, but may involve transcriptional control mechanisms.
In the HAP1 background, disruption of KLHL18 provides a powerful system to dissect its role in ubiquitin-mediated proteolysis within a leukemia-derived model. Given the near-haploid state, the polyclonal knockout population facilitates straightforward genotype-phenotype correlation and reduces the confounding effects of gene redundancy. This model is particularly suited to investigate the contribution of KLHL18 to cancer cell biology, including potential roles in cell proliferation, survival, and migration. Furthermore, the BCR-ABL-positive context allows researchers to explore functional interactions between KLHL18-dependent degradation pathways and oncogenic signaling networks relevant to hematological malignancies.
Typical research applications include ubiquitination assays using co-immunoprecipitation of ubiquitinated proteins, proteasomal degradation analysis, and cell proliferation assays such as MTT or BrdU incorporation. The KLHL18 knockout cells are also amenable to migration and invasion assays, flow cytometry-based cell cycle analysis, and transcriptomic profiling via RNA-seq. Additionally, mass spectrometry-based proteomics can be employed to identify novel KLHL18 substrates. This cell model supports drug target validation and screening for modulators of KLHL18 activity. For further technical details or assistance, please contact Ascent Research.
