The MTAP Knockout HCT 116 Cell Line is a CRISPR/Cas9-edited human knockout cell line designed to disrupt the MTAP gene, providing a stable loss-of-function model for investigating methylthioadenosine phosphorylase (MTAP) in colorectal carcinoma. Derived from the HCT 116 epithelial cell line, this product enables precise studies of MTAP-mediated pathways without transient gene silencing. The CRISPR/Cas9-mediated gene disruption ensures consistent genotype, supporting reproducible research in cancer metabolism and targeted therapy.
The HCT 116 host cell line originates from human colorectal carcinoma and exhibits microsatellite instability (MSI) due to a homozygous MLH1 mutation, with wild-type p53 expression. This well-characterized model is widely used for colorectal cancer studies, particularly for examining metabolic alterations and drug responses linked to genomic instability. Its robust growth and defined signaling make it suitable for detailed mechanistic and high-throughput analyses.
MTAP catalyzes the reversible phosphorolysis of methylthioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate, a critical step in the methionine salvage pathway that links polyamine metabolism to S-adenosylmethionine (SAM) recycling. MTAP deficiency leads to MTA accumulation, which inhibits protein arginine methyltransferase 5 (PRMT5) and spermidine synthase, creating a synthetic lethal dependency on methionine adenosyltransferase 2A (MAT2A). The MTAP gene is frequently co-deleted with CDKN2A on chromosome 9p21 and is transcriptionally regulated by MYC. These interactions place MTAP at the nexus of one-carbon metabolism, purine biosynthesis, and epigenetic regulation, with MTA, PRMT5, MAT2A, SAM, and spermidine serving as key molecular nodes.
In HCT 116 cells, MTAP knockout recapitulates metabolic vulnerabilities observed in MTAP-deleted tumors, including MTA buildup and altered polyamine flux. This model enables investigation of synthetic lethality strategies targeting PRMT5 or MAT2A within a defined genetic background of MSI and wild-type p53. The combination of MTAP loss and MSI may reveal interactions between metabolic stress and DNA repair, offering a relevant platform for preclinical drug evaluation in colorectal cancer.
This cell line is suitable for a range of applications, including synthetic lethality screens with PRMT5 inhibitors (e.g., GSK3326595), mass spectrometry-based quantification of MTA and SAM levels, and RNA-sequencing to assess transcriptomic changes. Standard validation involves western blotting for MTAP protein depletion and RT-qPCR for mRNA knockdown. Functional studies can utilize cell viability assays under PRMT5 or MAT2A inhibition, flow cytometry for apoptosis, and colony formation assays to measure long-term proliferation. The line also supports analyses of polyamine and methionine salvage pathways to dissect metabolic dependencies. For more information or to request a quote, contact Ascent Research.
