{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:14:52Z","timestamp":1785420892318,"version":"3.56.0"},"posted":{"date-parts":[[2026,1,18]]},"group-title":"Genetics","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T00:00:00Z","timestamp":1768694400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["Junior Investigator Award"],"award-info":[{"award-number":["Junior Investigator Award"]}],"id":[{"id":"https:\/\/ror.org\/01sx4nq39","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Research Grant Award"],"award-info":[{"award-number":["Research Grant Award"]}],"id":[{"id":"https:\/\/ror.org\/02gegq725","id-type":"ROR","asserted-by":"publisher"}]},{"name":"NIH NHGRI","award":["IGVF Program"],"award-info":[{"award-number":["IGVF Program"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Obtaining a precise genetic tuberous sclerosis diagnosis is a challenge as many missense\n                  <jats:italic>TSC2<\/jats:italic>\n                  variants are variants of uncertain significance (VUS). VUS in\n                  <jats:italic>TSC2<\/jats:italic>\n                  have been resolved by one-at-a-time functional assays, but these assays cannot scale to the 3,634\n                  <jats:italic>TSC2<\/jats:italic>\n                  missense VUS observed so far. To address this challenge, we used massively parallel sequencing to measure the steady-state abundance of almost 9,000\n                  <jats:italic>TSC2<\/jats:italic>\n                  missense variants and developed an mTOR pathway activity assay using genome editing and cell sorting to generate activity scores for 391 missense variants. 1,288 of 8,891 (14.49%) missense variants assayed had altered TSC2 abundance, and 69 of 391 (17.65%) missense variants assayed had altered mTOR pathway activity. Calibration and integration of these data into classification of variants identified in a clinical cohort putatively reclassified 212 of 276 (76.8%)\n                  <jats:italic>TSC2<\/jats:italic>\n                  missense VUS. These datasets will lead to improved genetic diagnosis of tuberous sclerosis with potential positive impacts on the clinical management of patients and their families.\n                <\/jats:p>","DOI":"10.64898\/2026.01.16.699909","type":"posted-content","created":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T04:20:13Z","timestamp":1768796413000},"source":"Crossref","is-referenced-by-count":2,"title":["An integrated, scaled approach to resolve TSC2 variants of uncertain significance"],"prefix":"10.64898","author":[{"given":"Carina G","family":"Biar","sequence":"first","affiliation":[{"name":"Ken and Ruth Davee Department of Neurology, Northwestern Feinberg School of Medicine, Chicago, Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyu R","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan D.","family":"Camp","sequence":"additional","affiliation":[{"name":"Center for Immunity and Immunotherapies, Seattle Children\u2019s Research Institute, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel L","family":"Holmes","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melinda K","family":"Wheelock","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sriram","family":"Pendyala","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abby V","family":"McGee","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, 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WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha","family":"Dawson","sequence":"additional","affiliation":[{"name":"Center for Immunity and Immunotherapies, Seattle Children\u2019s Research Institute, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Predrag","family":"Radivojac","sequence":"additional","affiliation":[{"name":"The Institute for Experiential AI, Northeastern University, Boston, MA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea M.","family":"Starita","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, WA"},{"name":"Brotman Baty Institute for Precision Medicine, Seattle, WA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gemma L","family":"Carvill","sequence":"additional","affiliation":[{"name":"Ken and Ruth Davee Department of Neurology, Northwestern Feinberg School of Medicine, Chicago, 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