The INTS10 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the INTS10 gene in the near-haploid human HAP1 cell line. This gene-edited model provides a loss-of-function genetic background for investigating the role of INTS10 in snRNA processing and transcription regulation. The polyclonal format eliminates the need for single-cell cloning, preserving population-level heterogeneity while enabling functional studies without the risk of clonal artifacts.
HAP1 cells are a fibroblast-like, near-haploid cell line derived from a male patient with chronic myeloid leukemia (CML). Their haploid karyotype facilitates straightforward gene knockout studies, as a single targeted disruption can lead to a complete absence of gene function. This makes HAP1 an ideal platform for genetic screens and detailed molecular analysis of genes involved in essential processes such as RNA metabolism and transcriptional control.
INTS10 encodes a core subunit of the Integrator complex, a multi-protein assembly that associates with RNA polymerase II (Pol II) to mediate 3?? end cleavage of nascent small nuclear RNAs (snRNAs) and regulate transcription elongation of protein-coding genes. Within this complex, INTS10 interacts directly with other integrator subunits, including INTS1, INTS4, INTS9, and INTS11, and contributes to the scaffolding that bridges the cleavage module to Pol II. Loss of INTS10 disrupts snRNA biogenesis, particularly of U1 and U2 snRNAs, and impairs Pol II transcription termination, leading to widespread effects on gene expression. Upstream regulators include Pol II and integrator assembly factors, while downstream targets encompass snRNA genes and a broad spectrum of Pol II-transcribed genes. This mechanistic integration positions INTS10 at a critical node in coupling transcription and RNA processing.
In the HAP1 background, knockout of INTS10 provides a stringent loss-of-function model that leverages the cell line??s near-haploid nature to ensure complete gene disruption. This is particularly valuable for studying the essential roles of integrator subunits, where hypomorphic alleles may obscure phenotypic consequences. The model enables dissection of INTS10??s contribution to snRNA processing and Pol II transcription, with direct relevance to neurodevelopmental disorders and cancer, where splicing and transcription defects are increasingly recognized as pathogenic drivers. The HAP1 system further facilitates high-throughput genetic screening to identify synthetic lethal interactions or modulators of RNA processing pathways.
Researchers can employ this INTS10 knockout polyclonal population in a variety of functional genomics and RNA biology applications. Western blotting and immunofluorescence confirm loss of INTS10 protein, while RT-qPCR and RNA-seq assays quantify changes in snRNA levels and transcriptome-wide alterations. Co-immunoprecipitation experiments enable assessment of Integrator complex integrity and Pol II association. This model is suitable for cancer biology studies investigating RNA processing dysregulation, as well as for screening small molecules or genetic interactions that rescue or exacerbate the knockout phenotype. For detailed experimental protocols or further assistance, please contact Ascent Research.
