The GPATCH11 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human knockout cell population designed for loss-of-function studies of the GPATCH11 gene. The polyclonal format provides a heterogeneous pool of cells with diverse CRISPR-mediated disruptions at the target locus, enabling robust population-level phenotypic analyses without clonal selection artifacts. This product is intended for biomedical researchers investigating gene function in an epithelial colorectal adenocarcinoma background.
HT29 cells are a widely used human colorectal adenocarcinoma cell line with epithelial morphology, originally derived from a 44-year-old female patient. These cells serve as a well-characterized model for intestinal epithelial biology and colorectal cancer, recapitulating key aspects of tumorigenesis including aberrant signaling, proliferation, and differentiation. Their genetic and phenotypic stability make them suitable for functional genomics studies.
GPATCH11 encodes a G-patch domain-containing protein that plays a regulatory role in pre-mRNA splicing, a critical process for transcriptomic diversity. The protein is known to interact with core spliceosome components such as SF3B1 and U2AF2, participating in the assembly and regulation of the spliceosome. Disruption of GPATCH11 is expected to impair spliceosomal function, leading to global alterations in alternative splicing patterns. This can affect downstream expression of genes involved in cell cycle control, apoptosis, and migration by shifting splicing isoform profiles of spliceosome-regulated transcripts. The knockout therefore creates a system to dissect splicing-dependent regulatory mechanisms.
In the colorectal cancer context, the HT29 GPATCH11 knockout model provides a powerful tool for investigating the role of splicing dysregulation in tumor cell biology. Since aberrant splicing is a hallmark of many cancers, including colorectal cancer, this model can be used to identify splicing vulnerabilities and evaluate therapeutic targeting of the spliceosome. The polyclonal population reflects the heterogeneity of editing outcomes, closely mimicking the genetic variability found in tumors, which is advantageous for studying drug responses and resistance mechanisms.
Research applications include detailed splicing analyses by RNA-seq and RT-qPCR for alternative splicing events, as well as functional assays such as MTS proliferation, apoptosis, and migration assays. The cells are also compatible with Western blotting to assess changes in protein isoform expression. This model is suitable for functional genomics screens to identify splicing-dependent growth regulators and for validating drug targets within the spliceosome pathway. For additional information or technical support, please contact Ascent Research.
