KHSRP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the HAP1 near-haploid human cell line, engineered to disrupt the KHSRP gene. This loss-of-function model enables investigation of KHSRP’s roles in mRNA decay and microRNA maturation. The polyclonal format, generated by CRISPR/Cas9-mediated gene disruption, enables population studies without clonal selection, maintaining heterogeneity. Researchers can employ this model to analyze KHSRP-dependent transcriptome changes, cytokine profiles, and signaling dynamics in a simplified genetic background.
The HAP1 cell line, derived from chronic myeloid leukemia KBM-7 cells, features a near-haploid karyotype. This haploid background permits unambiguous genotype-phenotype connections. HAP1 cells are highly amenable to CRISPR editing, exhibit robust transfection efficiency, and sustain vigorous growth, making them ideal for knockout screens and pathway analysis. Their genetic simplicity enhances the clarity of KHSRP knockout studies.
KHSRP binds AU-rich elements (AREs) in the 3?? UTRs of target mRNAs, facilitating their degradation via exosome recruitment. Validated targets include TNF-??, IL-6, IL-8, COX-2, and c-fos transcripts. Additionally, KHSRP promotes microRNA processing through interactions with the Drosha-DGCR8 microprocessor and Dicer, contributing to let-7 maturation. AKT-mediated phosphorylation and p38 MAPK activation modulate KHSRP, linking stress signals to gene regulation. KHSRP also interacts with 14-3-3 and tristetraprolin, integrating into the ARE-binding network.
Ablation of KHSRP in HAP1 cells disrupts ARE-mediated mRNA decay, stabilizing pro-inflammatory transcripts and enhancing NF-??B-dependent responses. This dysregulation amplifies NF-??B inflammatory responses and alters stress signaling, modeling immune dysregulation and cancer-related gene changes. Simultaneously, the loss of KHSRP impairs miRNA maturation, creating a dual-impact model for studying post-transcriptional dysregulation. The haploid state ensures penetrant effects, enabling precise dissection without allelic buffering.
This knockout model is suited for mRNA stability measurements via transcriptional chase assays, ARE-luciferase reporter analyses, and RNA-seq to identify KHSRP-regulated genes. MicroRNA expression analysis and RNA immunoprecipitation can clarify defects in miRNA biogenesis and binding targets. The cells support studies in inflammation biology, cancer research, and cellular stress responses, facilitating drug target validation and pathway interrogation. For further details and technical assistance, please contact Ascent Research.
