This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells with targeted disruption of the INTS4 gene. The polyclonal pool is generated by introducing Cas9 and a guide RNA specific to INTS4, followed by enrichment for loss-of-function mutations, yielding a heterogeneous cell population ideal for studying Integrator complex function without clonal bias. This knockout model enables robust interrogation of INTS4-dependent processes in a human cancer cell context.
HeLa cells, a human cervical adenocarcinoma line, are a cornerstone of biomedical research due to their immortalized nature and ease of genetic manipulation. They retain active RNA polymerase II machinery and are extensively used to study transcription regulation and oncogenic signaling. The INTS4 knockout in this background provides a relevant cancerous platform to investigate the role of the Integrator complex in gene expression control.
INTS4 encodes a critical subunit of the Integrator complex, which associates with RNA polymerase II to mediate 3′-end cleavage of nascent snRNAs (U1, U2) and regulate promoter-proximal pause-release. INTS4 interacts with multiple Integrator components (INTS1, INTS5, INTS7, INTS9?C12) and is regulated by factors such as CDK9 and NELF. Knockout of INTS4 abolishes Integrator endonuclease activity, leading to accumulation of unprocessed snRNAs and wide-ranging transcriptional abnormalities, highlighting its role in coordinating snRNA maturation and Pol II elongation.
In HeLa cells, loss of INTS4 disrupts Integrator complex assembly, resulting in defective snRNA processing and altered RNA polymerase II distribution. This triggers cell cycle dysregulation and impaired DNA damage responses, pathways commonly altered in cervical adenocarcinoma. The INTS4 knockout HeLa polyclonal cells thus serve as a valuable model to dissect how Integrator dysfunction fuels oncogenic phenotypes and to explore therapeutic vulnerabilities in cancers with transcriptional dependencies.
This knockout cell population is suited for diverse applications, including functional analysis of the Integrator complex, snRNA biogenesis studies, and transcription elongation assays. Standard techniques such as western blotting for INTS4 and Integrator subunits, RT-qPCR for snRNA processing, RNA-seq for transcriptome profiling, ChIP-qPCR for RNA polymerase II occupancy, and immunofluorescence for complex localization can be employed. The model also enables flow cytometry-based cell cycle analysis and drug target validation. For further details, contact Ascent Research.
