The KHSRP Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, designed for loss-of-function studies of the RNA-binding protein KHSRP (KH-type splicing regulatory protein). This gene-edited pool enables investigation of KHSRP??s role in post-transcriptional gene regulation without requiring clonal isolation, offering a heterogeneous population with targeted gene disruption.
The HT29 host cell line, isolated from a female colorectal adenocarcinoma, displays an adherent epithelial morphology and is extensively employed as a model system for intestinal epithelial biology. It recapitulates key aspects of intestinal barrier function, absorption, and colorectal tumorigenesis, making it a robust background for examining genes involved in mucosal homeostasis and cancer pathology.
KHSRP is an RNA-binding protein that predominantly promotes the decay of messenger RNAs containing AU-rich elements (AREs) in their 3?? untranslated regions. It functions by recruiting the exosome complex to target transcripts such as TNF??, c-Myc, IL-6, and p21, thereby modulating inflammatory and proliferative signaling. Upstream, KHSRP activity is regulated by p53-dependent transcriptional activation and post-translational modifications including AKT-mediated phosphorylation and MAPK pathway signals. In addition to its role in mRNA turnover, KHSRP participates in microRNA biogenesis by binding to primary microRNAs and enhancing their cleavage by the Drosha/DGCR8 microprocessor complex, with demonstrated interactions involving AUF1 and Argonaute 2 (Ago2).
Within the HT29 colorectal cancer model, ablation of KHSRP disrupts the normal processing of ARE-containing messages and microRNAs, providing a powerful tool to dissect the post-transcriptional networks that govern intestinal epithelial cell behavior. This knockout model is particularly relevant for studying how KHSRP-dependent regulation of targets like c-Myc and p21 impacts cell cycle progression, apoptosis, and motility, and how its interaction with inflammatory mediators such as TNF?? and IL-6 contributes to colitis-associated cancer phenotypes.
These polyclonal knockout cells are suited for a broad spectrum of molecular and cellular assays. Researchers can employ ARE-luciferase reporter systems to quantify KHSRP-mediated mRNA decay, RT-qPCR and western blotting to validate changes in target gene expression, RNA sequencing for transcriptome-wide analysis, and microRNA processing assays to assess biogenesis efficiency. Functional investigations can include cell proliferation, migration, and drug sensitivity screens to explore colorectal cancer progression and therapeutic responses. For technical inquiries, please contact Ascent Research.
