GPATCH3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population offering targeted disruption of the GPATCH3 gene. These cells are derived from the HAP1 human near-haploid cell line and harbor heterogeneous genetic lesions that abrogate GPATCH3 protein function. The polyclonal format avoids clonal selection artifacts, making it suitable for experiments requiring population-level phenotypes such as pooled genetic screens and bulk biochemical assays. This loss-of-function model enables systematic investigation of GPATCH3??s role in pre-mRNA splicing and RNA processing.
The HAP1 cell line is a near-haploid human male line originating from the KBM-7 chronic myeloid leukemia line. Its haploid nature allows recessive phenotypes to manifest after a single mutagenic event, greatly facilitating genetic screening. HAP1 cells maintain a stable haploid state in a large fraction of the population, providing a simplified genetic background for functional genomics studies. This characteristic makes HAP1 an ideal host for knockout models aimed at dissecting essential gene functions and signaling networks.
GPATCH3 encodes a G-patch domain-containing protein that acts as a cofactor for the RNA helicases DHX15 and DHX16 during pre-mRNA splicing. It directly interacts with these helicases and spliceosomal snRNPs to stimulate ATP-dependent conformational changes required for spliceosome assembly and catalysis. Through its G-patch domain, GPATCH3 facilitates helicase-driven remodeling of RNA-protein complexes, ensuring accurate intron removal and exon ligation. Disruption of GPATCH3 therefore impairs spliceosome dynamics, leading to widespread splicing defects such as intron retention and exon skipping, which compromise gene expression and cellular fitness.
In the HAP1 near-haploid background, GPATCH3 knockout eliminates the wild-type allele, removing any possibility of compensation and revealing direct loss-of-function phenotypes. This clean genetic model is particularly valuable for studying splicing factor dependencies and for identifying genetic interactions via synthetic lethality screens. The combination of haploid genetics and splicing disruption provides a powerful system to explore the vulnerability of cancer cells to spliceosome perturbations, given the known dependence of many tumors on efficient RNA processing.
These knockout cells support a range of research applications, including RNA-seq for transcriptome-wide splicing analysis, RT-PCR for validation of specific splice isoforms, and co-immunoprecipitation to confirm interactions with DHX15 or DHX16. Additional assays such as western blotting for spliceosome components and cell viability measurements can assess the functional impact of GPATCH3 loss. The polyclonal knockout pool is also well-suited for CRISPR-based modifier screens seeking regulators of splicing fidelity. For technical inquiries or custom requests, contact Ascent Research.
