{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T06:52:10Z","timestamp":1791615130468,"version":"4.3.4"},"reference-count":53,"publisher":"National Academy of Sciences","issue":"3","license":[{"start":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T00:00:00Z","timestamp":1768521600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004622","name":"Dutch Cancer Society","doi-asserted-by":"crossref","award":["10645"],"award-info":[{"award-number":["10645"]}],"id":[{"id":"10.13039\/501100004622","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004622","name":"Dutch Cancer Society","doi-asserted-by":"crossref","award":["14469"],"award-info":[{"award-number":["14469"]}],"id":[{"id":"10.13039\/501100004622","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100003246","name":"Dutch Research Council","doi-asserted-by":"crossref","award":["TTW 1488"],"award-info":[{"award-number":["TTW 1488"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,1,20]]},"abstract":"<jats:p>\n                    Lynch syndrome (LS) is a genetic predisposition to mainly colorectal and endometrial cancer due to heterozygous disruptive germline mutations in the DNA mismatch-repair (MMR) genes\n                    <jats:italic toggle=\"yes\">MSH2<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">MSH6<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">MLH1,<\/jats:italic>\n                    or\n                    <jats:italic toggle=\"yes\">PMS2<\/jats:italic>\n                    . Beyond clearly pathogenic mutations, germline sequencing often reveals variants of uncertain significance (VUS), predominantly single base-pair alterations in coding or noncoding regions. These uncertain variants obstruct LS diagnosis, hampering personalized surveillance. To address this challenge, we developed a highly accurate functional assay that interrogates VUS pathogenicity in human cells. Building on a mouse-based cellular assay, we adapted oligonucleotide-directed mutation screening (ODMS) for human cells and introduced a refined approach named \u201ccoselection ODMS.\u201d To ensure physiological expression, the variant is introduced into the endogenous MMR gene by replication-coupled gene editing. Coselection ODMS demonstrated 100% accuracy in classifying 50 benign and 86 pathogenic variants spanning coding and noncoding regions in all four MMR genes. Among 109 patient-derived VUS, 51 were identified as deleterious for MMR function. Importantly, coselection ODMS delivered 100% concordant results in a clinical diagnostic laboratory. With &gt;93% sensitivity and &gt;92% specificity, coselection ODMS provides a highly reliable functional assay in the diagnosis of enigmatic LS variants, enabling risk assessment and personalized surveillance or treatment for affected families.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2517801123","type":"journal-article","created":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T17:06:13Z","timestamp":1768583173000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":1,"title":["An accurate cellular assay to determine pathogenicity of coding and noncoding variants in Lynch syndrome genes"],"prefix":"10.1073","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1829-2403","authenticated-orcid":false,"given":"Iris E.","family":"Glykofridis","sequence":"first","affiliation":[{"name":"Division of Tumor Biology and Immunology, The Netherlands Cancer Institute","place":["Amsterdam, The 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