The KHSRP Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the KHSRP gene has been disrupted, creating a loss-of-function model for studying post-transcriptional gene regulation. This polyclonal pool contains a heterogeneous collection of edited HEK293T cells, each carrying distinct CRISPR/Cas9-mediated gene disruptions within KHSRP, enabling robust representation of knockout phenotypes without single-cell cloning. The population format is ideal for researchers requiring immediate access to a functional knockout model while preserving biological variability inherent to polyclonal cultures.
The host cell line, HEK293T, is a clonal derivative of the human embryonic kidney HEK293 cell line, constitutively expressing the SV40 large T antigen. These adherent epithelial cells are widely employed in biomedical research due to their high transfectability, rapid growth, and suitability for recombinant protein expression, viral packaging, and transient transfection. Their well-characterized genetic background and compatibility with various functional assays make them a versatile platform for investigating gene function, particularly in the context of signaling and RNA biology.
KHSRP encodes an RNA-binding protein that orchestrates two critical facets of post-transcriptional control: mRNA decay and microRNA (miRNA) biogenesis. Mechanistically, KHSRP binds AU-rich elements (AREs) in the 3?? untranslated regions of target mRNAs, recruiting the exosome complex (including subunits such as EXOSC3) to accelerate transcript degradation. This activity downregulates key proliferative and inflammatory mediators, including VEGF, MYC, CCND1, IL-6, and TNF-??. Concurrently, KHSRP facilitates miRNA maturation by interacting with the terminal loop of primary miRNA transcripts and recruiting the Drosha-DGCR8 Microprocessor complex, thereby promoting processing of miRNAs like let-7 and miR-21. Upstream, KHSRP is regulated by TGF-?? receptor/SMAD signaling, Wnt/??-catenin pathways, MAP kinases (ERK, p38), and NF-??B, positioning it as a nodal integrator of extracellular cues. Its interactome further includes UPF1, PARN, hnRNP K, AUF1, and VHL, underscoring its multifaceted roles.
In HEK293T cells, which lack tissue-specific signaling nuances, KHSRP knockout allows dissection of its fundamental regulatory activities in a simplified, experimentally tractable system. The presence of SV40 large T antigen, which neutralizes p53 and pRb tumor suppressors, creates a permissive background for analyzing KHSRP??s impact on cell cycle progression and senescence, given its downstream target p21/CDKN1A. This model thus provides a unique window into how KHSRP modulates ARE-mediated decay and miRNA processing without confounding developmental or lineage-specific variables, making it suitable for biochemical reconstitution and mechanistic studies.
This knockout cell population supports a broad array of experimental workflows. Researchers can employ RT-qPCR, RNA sequencing, and luciferase reporter assays to quantify mRNA stability and ARE-dependent decay, while processing assays and small RNA sequencing reveal alterations in miRNA biogenesis. Western blotting and co-immunoprecipitation enable analysis of KHSRP interactors and downstream protein expression. Functionally, the model facilitates flow cytometry-based cell cycle analysis, migration and invasion assays, and investigations into TGF-??/Wnt-driven signaling in cancer and inflammation. For further technical inquiries or ordering information, please contact Ascent Research.
