The KHSRP Knockout HeLa Polyclonal Cells product comprises a polyclonal population of HeLa cells with CRISPR/Cas9-mediated disruption of the KHSRP gene, encoding the KH-type splicing regulatory protein. This polyclonal knockout model provides a heterogeneous loss-of-function background for investigating KHSRP-dependent post-transcriptional gene regulation. The product is supplied as a live cell population suitable for immediate expansion in culture, enabling robust experimental replication. By utilizing a polyclonal format, the model preserves genetic diversity, mitigating the effects of clonal selection and enhancing physiological representativeness in functional studies.
The host HeLa cell line is an extensively characterized human cervical adenocarcinoma model, originally derived from Henrietta Lacks, and is HPV18-positive. HeLa cells are widely employed in cancer biology, signal transduction, and drug discovery research due to their robust growth and well-documented molecular profiles. The presence of integrated HPV18 oncoproteins E6 and E7 provides a relevant context for studying interactions between viral transformation mechanisms and host regulatory pathways, making HeLa an ideal platform for exploring the functions of RNA-binding proteins like KHSRP.
KHSRP is a multifunctional RNA-binding protein that post-transcriptionally regulates gene expression. It binds AU-rich elements in mRNAs such as c-FOS, TNF??, and MYC, promoting their exosome-mediated decay, and interacts with Drosha and Dicer to process miRNA precursors like miR-155 and let-7. Upstream kinases including p38 MAPK, ERK, and AKT, as well as interleukin-1, modulate KHSRP activity, while KHSRP cooperates with EXOSC3, UPF1, and hnRNP A1 in RNA metabolic complexes. This positions KHSRP at the intersection of mRNA turnover and miRNA biogenesis, integrating signals from diverse cellular pathways.
In HeLa cells, disruption of KHSRP can profoundly alter the post-transcriptional landscape, affecting the expression of oncogenes and inflammatory mediators within an HPV18-positive cervical adenocarcinoma context. The deregulation of c-FOS and MYC stability, TNF?? production, and let-7 miRNA levels may contribute to understanding how KHSRP loss impacts transformation-related pathways. Thus, this polyclonal knockout model enables investigation of KHSRP-dependent regulatory mechanisms in a well-established cancer cell system.
Researchers can employ this product in a variety of experimental contexts, including functional genomics, RNA biology, cancer cell signaling, and miRNA biogenesis studies. Representative assays include RT-qPCR for measuring target mRNA and miRNA levels, western blotting for assessing protein expression changes, RNA immunoprecipitation for analyzing KHSRP?CRNA interactions, and mRNA stability assays to evaluate decay kinetics. Additionally, miRNA profiling can elucidate global changes in miRNA expression networks following KHSRP loss. For additional information or technical support, please contact Ascent Research.
