GPATCH1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T line. This product consists of a heterogeneous pool of cells with targeted disruptions in the GPATCH1 gene, enabling loss-of-function studies without clonal selection. The polyclonal format is suitable for bulk assays, functional screens, and analyses where editing diversity captures the range of knockout phenotypes. The knockout is achieved via CRISPR/Cas9-mediated gene disruption, resulting in a population-level ablation of GPATCH1 function.
HEK293T cells are a widely used human embryonic kidney cell line stably expressing the SV40 large T antigen, which supports episomal plasmid replication and enhances protein expression. Their high transfection efficiency and robust growth make them a preferred host for gene editing, lentivirus production, and recombinant protein expression. They retain intact RNA processing pathways, providing a physiologically relevant background for investigating splicing factors. The knockout model in this background enables precise dissection of GPATCH1 function in a tractable human cell system.
GPATCH1 contains a G-patch domain and is implicated in pre-mRNA splicing via its association with the spliceosome. It interacts with core spliceosomal components, including the U5 snRNP proteins SNRNP200 and PRPF8, and the U2 snRNP factor SF3B1. These interactions position GPATCH1 within the catalytic core, where it may facilitate conformational rearrangements during splicing. Loss of GPATCH1 can disrupt spliceosome assembly or function, leading to global splicing alterations and potential effects on ribosome biogenesis. Dysregulation of splicing is linked to oncogenesis, particularly in gastric cancer and glioma, making this knockout model relevant for cancer research.
In HEK293T cells, GPATCH1 knockout provides a platform to study splicing-dependent phenotypes in a fast-growing, epithelial context. The cells?? high transfectability allows for complementation with wild-type or mutant GPATCH1 constructs, enabling structure-function analyses. They can be used in co-immunoprecipitation experiments to probe spliceosome integrity, or in RNA-seq studies to map splicing changes genome-wide. This model also facilitates investigation of potential synthetic lethal interactions with splicing inhibitors, offering a tool for drug target validation.
Researchers can employ these polyclonal knockout cells to investigate mRNA splicing mechanisms, RNA processing regulation, and cancer cell biology. Representative assays include Western blotting for GPATCH1 validation, RT-qPCR and RNA-seq to analyze splicing variants, co-immunoprecipitation to assess SNRNP200 or PRPF8 interactions, and immunofluorescence to examine subcellular localization. Cell proliferation and drug sensitivity assays may reveal phenotypes relevant to gastric cancer and glioma research. For ordering and technical inquiries, please contact Ascent Research.
