PMS2 Knockout Cell Line (HEK293)

The choice depends on whether you are studying PMS2's role as the dominant MutLα partner of MLH1 in mismatch repair or modeling Lynch syndrome and constitutional mismatch repair deficiency (CMMRD) syndrome. The Knockout line is the standard tool for asking whether PMS2 is required for mismatch repair — PMS2 partners with MLH1 to form MutLα, the principal MutL-family complex in human MMR, providing endonuclease activity that introduces strand-specific nicks during repair. Overexpression is useful for studying PMS2 in heterologous expression contexts or for testing disease-associated mutations. For mismatch repair research, the EDITGENE PMS2 Knockout in HEK293 is a workhorse mechanistic platform — HEK293 has been extensively used for MMR biochemistry and structure-function studies. PMS2 mutations cause Lynch syndrome (HNPCC) and, when biallelic, constitutional mismatch repair deficiency (CMMRD) — disease variant rescue enables genotype-function studies. Rescue with wild-type or endonuclease-dead (E705K) PMS2 enables comprehensive structure-function studies. The knockout is valuable for studying PMS2-deficient cancer immunotherapy response — MMR-deficient tumors have high mutation burden and increased sensitivity to immune checkpoint inhibitors.

Primary applications: • Mismatch repair activity: in vitro MMR reconstitution assays using nuclear extracts and defined heteroduplex substrates to quantify MMR proficiency in the absence of PMS2. • Microsatellite instability (MSI): repeat tract stability analysis (e.g., BAT-25, BAT-26) given MSI-high phenotype of MMR-deficient cells. • Lynch/CMMRD modeling: rescue with patient-derived PMS2 mutations for genotype-function studies. • Mutational signature analysis: whole-exome sequencing to characterize MMR-deficient mutational signatures (SBS6, SBS15, SBS20, SBS26). EDITGENE recommends this model for researchers investigating DNA mismatch repair, Lynch syndrome mechanisms, and MMR deficiency-associated cancer immunotherapy response.

Yes. PMS2 rescue experiments are well-established for MMR research: • Construct design: use a codon-modified PMS2 sequence with a small C-terminal tag (FLAG, HA). PMS2 has N-terminal ATPase domain, central CTH-binding region, and C-terminal endonuclease domain (DQHA-X(2)-E-X(4)-E motif) — preserve all elements. • Endonuclease-dead rescue: the E705K mutation in the conserved endonuclease motif abolishes nicking activity and is the standard specificity control. • MLH1 interaction: rescue interpretation considers MLH1 expression — PMS2 requires MLH1 for stability and MutLα formation. • Disease mutation rescue: Lynch/CMMRD-associated PMS2 mutations enable comprehensive disease genotype-function studies. • Functional readout: rescue should restore MMR activity (in vitro reconstitution), microsatellite stability, and 6-thioguanine sensitivity. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.