A calibrated cell-based functional assay to aid classification of MLH1 DNA mismatch repair gene variants.

ABSTRACT: Functional assays provide important evidence for classifying the disease significance of germline variants in DNA mismatch repair genes. Numerous laboratories, including our own, have developed functional assays to study mismatch repair gene variants. However, previous assays are limited due to the model system employed, the manner of gene expression, or the environment in which function is assessed. Here, we developed a human cell-based approach for testing the function of variants of uncertain significance (VUS) in the MLH1 gene. Using clustered regularly interspaced short palindromic repeats gene editing, we knocked in MLH1 VUS into the endogenous MLH1 loci in human embryonic stem cells. We examined their impact on RNA and protein, including their ability to prevent microsatellite insta

SUBMITTER: Rath A 

PROVIDER: S-EPMC9772141 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature