The INTS4 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population designed for loss-of-function studies of INTS4. This product provides a heterogeneous pool of HEK293T cells harboring targeted disruptions in the INTS4 gene, enabling researchers to investigate the functional consequences of Integrator complex subunit 4 depletion. As a polyclonal knockout model, it avoids clonal artifacts and offers a population-level view of gene disruption effects, suitable for experiments where a mixed-genotype pool is beneficial.
The HEK293T host cell line is a derivative of human embryonic kidney 293 cells, stably expressing the SV40 large T antigen. This modification facilitates episomal replication of plasmids containing the SV40 origin, resulting in high transfection efficiency and robust protein expression. Widely used in recombinant protein production and viral packaging, HEK293T cells provide a versatile and well-characterized background for studying gene function, particularly in the context of transcription and RNA processing.
INTS4 encodes a core subunit of the Integrator complex, a multi-protein assembly that associates with RNA Polymerase II to mediate 3?? end processing of small nuclear RNAs (snRNAs) and enhancer RNAs. Mechanistically, INTS4 functions within the Integrator complex alongside INTS1, INTS2, and CPSF73 to couple transcription termination with RNA maturation. It directly interacts with RNA Pol II and is essential for the accurate processing of downstream targets such as U1 snRNA, U2 snRNA, and the MYC proto-oncogene, facilitating proper termination of MYC transcription. Disruption of INTS4 impairs Integrator activity, leading to defective snRNA biogenesis and aberrant transcriptional regulation.
In the HEK293T background, INTS4 knockout cells serve as a tractable system to dissect Integrator complex functions. This model enables the study of snRNA processing and transcription termination mechanisms in a cell type with high transfection competence, facilitating both transient and stable complementation experiments. The dysregulation of MYC and other targets provides a platform to explore connections between Integrator dysfunction and oncogenic processes, while the polyclonal nature of the population allows for assessment of phenotypic variability and essential gene function without single-clone bias.
Typical applications include snRNA processing studies via RT-qPCR and RNA-seq, transcription regulation assays using ChIP-qPCR, and investigation of protein interactions through co-immunoprecipitation and western blotting. The model is also suited for cancer cell biology research, including cell proliferation assays and flow cytometry-based phenotypic analysis, and for modeling neurodevelopmental disorders linked to Integrator dysfunction. These polyclonal knockout cells support functional genomics screens and pathway analysis with relevant controls. For further information or to discuss customized experimental solutions, please contact Ascent Research.
