The GPATCH2 Knockout HEK293T Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of HEK293T cells, engineered for loss-of-function studies of the GPATCH2 gene. This heterogeneous knockout pool facilitates investigation of gene function without clonal selection biases, enabling robust assessment of splicing-dependent phenotypes. The polyclonal format is particularly suited for assays where population-level responses are prioritized, ensuring representation of multiple editing events across the cell population. This model provides a versatile platform for studying GPATCH2-mediated processes in a human cellular context.
HEK293T cells, derived from human embryonic kidney epithelium, are immortalized with sheared adenovirus 5 DNA and stably express the SV40 large T antigen. This antigen allows episomal replication of plasmids containing the SV40 origin, making the cell line exceptionally efficient for transient protein expression and viral vector production. The HEK293T background is widely adopted in molecular and cellular biology for its robust growth characteristics, high transfection efficiency, and well-characterized transcriptome, providing a standardized system for genome engineering and functional assays.
GPATCH2 encodes a nuclear G-patch domain-containing protein implicated in pre-mRNA splicing and nucleic acid metabolism. It interacts with core spliceosomal components, including CTNNBL1 and RBM17, and the KIAA1429 protein, within the spliceosome and PRP19 complex. These interactions suggest a regulatory role in mRNA maturation, influencing gene expression outputs. GPATCH2 expression is subject to transcriptional regulation and cell cycle-dependent control, and it modulates the splicing of target pre-mRNAs, thus affecting downstream gene expression programs. Aberrations in these processes link GPATCH2 to chromosome 6q11-q14 deletion syndrome and potential roles in cancer.
In the HEK293T background, disruption of GPATCH2 offers a tractable system to dissect splicing mechanisms and their impact on cellular physiology. The cell line??s high expression capacity and ease of transfection complement the knockout, enabling detailed biochemical and functional analyses. Researchers can examine how loss of GPATCH2 alters spliceosome assembly and the processing of specific pre-mRNA substrates, leveraging the well-annotated transcriptome of HEK293T cells. This model also supports studies into the intersection of splicing regulation and cancer cell biology, given the gene??s potential oncogenic links.
Typical applications include RNA-seq to profile alternative splicing changes, co-immunoprecipitation with CTNNBL1 or RBM17 to assess protein interactions, and RT-qPCR to quantify isoform expression. Western blotting confirms protein-level disruption, while proliferation and cell cycle assays evaluate phenotypic consequences of GPATCH2 loss. These tools support functional genomics, drug target exploration, and splicing mechanism investigations. For additional details on this product or related services, please contact Ascent Research.
