The KLHL7 Knockout HAP1 Polyclonal Cells represent a polyclonal knockout cell population derived from the HAP1 cell line following CRISPR/Cas9-mediated disruption of the human KLHL7 gene. This product provides a robust loss-of-function model for dissecting KLHL7-dependent cellular processes without introducing defined clonal genotypes. By ablating KLHL7 expression, researchers can interrogate the functional consequences of impaired CRL5 E3 ligase activity in a genetically tractable, near-haploid background.
HAP1 cells originate from the KBM-7 chronic myelogenous leukemia (CML) line and maintain a near-haploid karyotype with disomy for chromosome 8. They are BCR-ABL positive, semi-adherent, and exhibit fibroblast-like morphology. This unique genetic configuration facilitates high-efficiency CRISPR editing and unambiguous genotype?Cphenotype correlations, making HAP1 an ideal host for genetic screens and signaling studies in a hematological malignancy context.
KLHL7 encodes a substrate-specific adaptor that assembles with Cullin5 (CUL5), Rbx2, Elongin B (TCEB1), and Elongin C (TCEB2) to form a functional CRL5 E3 ubiquitin ligase complex. Through its Kelch-repeat domain, KLHL7 recruits substrates such as REF1 (RBMX) and I??B?? for polyubiquitination, targeting them for 26S proteasomal degradation. This activity is regulated by NF-??B?Cdependent transcriptional control, neddylation of CUL5, and potentially CDK5-mediated phosphorylation. Downstream consequences include modulation of NF-??B signaling, apoptotic progression, and cell cycle transition, positioning KLHL7 as a critical node in ubiquitin-mediated proteolysis.
In the HAP1 cell context, KLHL7 knockout permits a focused analysis of CRL5-dependent ubiquitination events and their impact on leukemogenic pathways. Given the BCR-ABL?Cdriven background, this model is particularly suited to examine crosstalk between tyrosine kinase signaling and protein degradation networks. The haploid genome simplifies detection of functional vulnerabilities, enabling high-confidence identification of synthetic lethal interactions or drug sensitivity modulators relevant to myeloid malignancies.
This polyclonal knockout pool is designed for diverse applications, including mechanistic studies of ubiquitin-proteasome system components, CRL5 E3 ligase target identification, and validation of substrates like REF1 and I??B??. Standard assays include Western blotting, ubiquitination assays, NF-??B luciferase reporters, caspase activity measurements, cell cycle flow cytometry, co-immunoprecipitation, and proteasome activity assays. The product also supports retinitis pigmentosa disease modeling (autosomal dominant RP42) and high-throughput haploid genetic screens. For further details or to customize your knockout cell needs, please contact Ascent Research.
