The GRSF1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T cell line, featuring targeted disruption of the GRSF1 gene. This product consists of a heterogeneous mixture of cells with diverse CRISPR-induced edits at the GRSF1 locus, yielding a loss-of-function population suitable for studies where gene inactivation at the pool level is desirable. The polyclonal format bypasses clonal isolation, providing a robust, ready-to-use model that maintains the established growth characteristics and experimental versatility of the parental line.
HEK293T is a human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, enabling high-level episomal replication of plasmids bearing the SV40 origin. This feature, coupled with exceptional transfectability and protein production capacity, has made HEK293T a backbone for recombinant protein expression, lentiviral/retroviral packaging, and functional genomics. Its epithelial lineage and active translational apparatus offer a physiologically relevant setting for dissecting RNA regulatory pathways and mitochondrial gene expression.
GRSF1 (G-rich RNA sequence binding factor 1) is an RNA-binding protein that recognizes G-rich elements and regulates the stability and translation of critical targets, including mitochondrial-encoded COX1, COX2, and ND1 transcripts and viral IRES-containing RNAs. It localizes to stress granules and mitochondria, interacting with RNA-binding factors hnRNP A1, TIA-1, TIAR, and components of the mitochondrial ribosome and PPR proteins. GRSF1 is modulated by mitochondrial stress, type I interferons, and viral infection, and operates within the mitochondrial gene expression cascade alongside TFAM and POLRMT, and in antiviral responses through the RIG-I, MDA5, and IRF3 signaling axis, while also collaborating with eIF4G and PTB to regulate IRES-dependent translation.
In HEK293T cells, GRSF1 knockout enables dissection of the links among mitochondrial gene regulation, RNA metabolism, and innate immunity. This model permits direct investigation of GRSF1??s role in stress granule dynamics, mitochondrial mRNA translation, and antiviral signaling via RIG-I/MDA5/IRF3. The host??s proficiency in supporting viral replication makes it ideal for studying how GRSF1-dependent RNA processing affects infection susceptibility.
Typical applications include Western blotting and RT-qPCR for target validation, RNA-seq for transcriptome profiling, and immunofluorescence for subcellular localization. Functional assays such as Seahorse metabolic analysis and polysome profiling evaluate mitochondrial respiration and translational control of mitochondrial mRNAs. Viral infection assays combined with expression analysis dissect GRSF1-dependent antiviral responses. The polyclonal knockout population is also suitable for pooled screening and functional genomics. For further details or custom engineering, contact Ascent Research.
