The ISOC2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ISOC2 gene in the human colorectal adenocarcinoma cell line HT29. This polyclonal population consists of a heterogeneous pool of cells carrying CRISPR/Cas9-mediated gene disruptions at the ISOC2 locus, providing a loss-of-function model for studying the biological role of ISOC2. The use of a polyclonal format offers advantages in capturing diverse genetic perturbation profiles, which can be valuable for population-level analyses of gene function in cancer and metabolic research.
The HT29 cell line was originally isolated from a primary colorectal adenocarcinoma of a 44-year-old female patient. These adherent epithelial-like cells serve as a well-established model for the intestinal epithelial barrier and colorectal cancer biology. HT29 cells are widely utilized in studies of epithelial polarization, tight junction formation, and cancer cell metabolism. Their human origin and retention of key colorectal cancer features make them a relevant host for investigating genes involved in mitochondrial metabolism and disease progression.
The ISOC2 gene encodes a protein containing a predicted hydrolase domain and is likely localized to mitochondria, suggesting a role in mitochondrial metabolite processing. Although its precise molecular interaction network remains poorly characterized, bioinformatic analyses indicate that ISOC2 may function as a mitochondrial hydrolase, potentially participating in the hydrolysis of metabolic intermediates. No specific upstream regulators, downstream targets, or interacting partners have been clearly defined. However, its hydrolase activity implies involvement in pathways such as mitochondrial metabolism and hydrolase-mediated reactions, and loss of ISOC2 may indirectly influence mitochondrial homeostasis and cellular stress responses.
In the context of HT29 colorectal cancer cells, knockout of ISOC2 offers a valuable model to explore how a putative mitochondrial hydrolase influences cancer cell metabolism. HT29 cells exhibit active mitochondrial oxidative phosphorylation and glycolysis, and ISOC2 disruption could perturb metabolic flux, energy production, or redox balance. This model may help uncover metabolic vulnerabilities specific to colorectal cancer and assess the impact of mitochondrial enzyme deficiencies on tumor cell growth and survival. It is particularly suited for examining the interplay between mitochondrial function and the metabolic adaptations of cancer cells.
This ISOC2 knockout polyclonal cell product is suitable for a range of functional and molecular assays. Researchers can confirm ISOC2 disruption using Western blotting, RT-qPCR, and Sanger sequencing. Metabolic consequences can be evaluated with Seahorse metabolic flux analysis to measure oxygen consumption and extracellular acidification, while cell viability and apoptosis can be assessed using MTT and annexin V assays, respectively. LC-MS metabolomics enables in-depth profiling of metabolite changes. Applications extend to colorectal cancer metabolism research, mitochondrial biology, and functional genomics screens. For additional information, please contact Ascent Research.
