{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T10:01:38Z","timestamp":1753869698167,"version":"3.41.2"},"reference-count":22,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2020,7,31]],"date-time":"2020-07-31T00:00:00Z","timestamp":1596153600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81271364"],"award-info":[{"award-number":["81271364"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molec Gen &amp;amp; Gen Med"],"published-print":{"date-parts":[[2020,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Tuberous sclerosis complex (TSC), belongs to autosomal dominant genetic disorder, which affects multiple organ systems in the body, including the skin, brain, lungs, kidneys, liver, and eyes. Mutations in <jats:italic>TSC1<\/jats:italic> or <jats:italic>TSC2<\/jats:italic> was proved to be associated with these conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Gene\u2010panel Sequence of NGS was used to detect the mutation in a Chinese family. The research further investigates whether aberrant splicing and nonsense\u2010mediated mRNA degradation (NMD) could serve as a mechanism cause by <jats:italic>TSC1<\/jats:italic> mutation. MINI\u2010Gene assay apply by pcMINI\u2010<jats:italic>TSC1<\/jats:italic>wt\/mut plasmids delivered in HeLa and 293T cell lines. Recombinant plasmids expressing wild\u2010type and mutant\u2010type EGFP\u2010<jats:italic>TSC1<\/jats:italic> were constructed and transiently transfected into human embryonic kidney cells 293T by lipofectamine. Real\u2010time PCR and Western Blot were performed to analyze the expression of mRNAs and proteins of EGFP\u2010<jats:italic>TSC1<\/jats:italic> and NMD factor UPF1.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The gene test verified a novel heterozygous <jats:italic>TSC1<\/jats:italic> frameshift mutation (<jats:italic>TSC1<\/jats:italic> c.1550_1551del) in the proband and her mother. From MINI\u2010Gene assay, the agarose gel showed that both the mutant and wild\u2010type mRNA possess two main bands, indicating two splicing modes, named band A and B, respectively. The mutation c.1550_1551del has not produced new splicing site, but there is a selective splicing in varying degree significantly after mutation. On the contrary, function validation assay showed that cells transfected with the mutant <jats:italic>TSC1<\/jats:italic> plasmids expressed significantly lower <jats:italic>TSC1<\/jats:italic> in mRNAs and proteins levels, compared with the wild\u2010type <jats:italic>TSC1<\/jats:italic> plasmid transfection. A translation inhibitor cycloheximide and small interfering RNA of UPF1 (siRNA\u2010UPF1) increased mRNA or protein expression of <jats:italic>TSC1<\/jats:italic> significantly in cells transfected with the mutant plasmids.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our study demonstrated that the novel <jats:italic>TSC1<\/jats:italic> frameshift mutation (<jats:italic>TSC1<\/jats:italic> c.1550_1551del) trigger aberrant splicing and NMD simultaneously, causing decrease of hamartin, then, leading to tuberous sclerosis complex formation.<\/jats:p><\/jats:sec>","DOI":"10.1002\/mgg3.1410","type":"journal-article","created":{"date-parts":[[2020,7,31]],"date-time":"2020-07-31T11:07:27Z","timestamp":1596193647000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A novel TSC1 frameshift mutation c.1550_1551del causes tuberous sclerosis complex by aberrant splicing and nonsense\u2010mediated mRNA degradation (NMD) simultaneously in a Chinese family"],"prefix":"10.1002","volume":"8","author":[{"given":"Cong","family":"Qiu","sequence":"first","affiliation":[{"name":"Department of Neurology Renmin Hospital of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chengyan","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Neurology Renmin Hospital of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyun","family":"Tong","sequence":"additional","affiliation":[{"name":"Department of Neurology Renmin Hospital of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luoyang","family":"Dai","sequence":"additional","affiliation":[{"name":"Department of Neurology Renmin Hospital of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenda","family":"Liu","sequence":"additional","affiliation":[{"name":"Demonstration Center for Experimental Basic Medicine Education of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yulie","family":"Xie","sequence":"additional","affiliation":[{"name":"Demonstration Center for Experimental Basic Medicine Education of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qimei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Central hospital of Yichang City The First Clinical Medical College of Three Gorges University Yichang Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5631-8139","authenticated-orcid":false,"given":"Guohua","family":"Yang","sequence":"additional","affiliation":[{"name":"Demonstration Center for Experimental Basic Medicine Education of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Neurology Renmin Hospital of Wuhan University Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,7,31]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2006.83"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1212\/WNL.0b013e3181e04325"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1086\/316951"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2012.06.009"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12041\u2010018\u20100927\u20107"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng0294\u2010193"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1089\/gtmb.2019.0094"},{"issue":"2","key":"e_1_2_9_9_1","first-page":"400","article-title":"Allelic Loss is frequent in tuberous sclerosis kidney lesions but rare in brain lesions","volume":"59","author":"Henske E. 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