{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T05:43:44Z","timestamp":1775195024298,"version":"3.50.1"},"reference-count":22,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2020,8,10]],"date-time":"2020-08-10T00:00:00Z","timestamp":1597017600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Annals of Human Genetics"],"published-print":{"date-parts":[[2020,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Variants perturbing the normal splicing of pre\u2010mRNA can lead to human diseases. The splice\u2010altering effect and eventual consequence on gene function was sometimes uncertain and hinders a definitive molecular diagnosis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The impact of four rare intronic variants on splicing was analyzed through reverse transcription \u2010 polymerase chain reaction (RT\u2010PCR) analysis of mRNA derived from the peripheral blood of patients. The results were compared with in\u2010silico prediction. Potential implication on molecular diagnosis was discussed.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Four rare intronic variants of <jats:italic>SLC9A6<\/jats:italic>, <jats:italic>DLG3<\/jats:italic>, <jats:italic>GAA<\/jats:italic>, and <jats:italic>OCRL<\/jats:italic> were identified in patients with suspected disorders, respectively. Although these four variants were all predicted to alter splicing by in\u2010silico tools, RT\u2010PCR analysis of mRNA derived from peripheral blood showed these variants affected splicing in different ways: c.899+3_899+6del of <jats:italic>SLC9A6<\/jats:italic> resulted in one\u2010exon skipping and an out\u2010of\u2010frame transcript; c.905\u20102A\u00a0&gt;\u00a0G of <jats:italic>DLG3<\/jats:italic> resulted in a mix of in\u2010frame transcripts; c.1195\u201011T\u00a0&gt;\u00a0A of <jats:italic>GAA<\/jats:italic> resulted in the in\u2010frame insertion of nine nucleotides; c.723\u20102A\u00a0&gt;\u00a0C of <jats:italic>OCRL<\/jats:italic> resulted in one\u2010exon skipping and in\u2010frame deletion of 102 nucleotides. The consequence revealed by mRNA analysis is essential for accurate interpretation of pathogenicity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Four intronic variants all caused aberrant mRNA splicing. For intronic variants with uncertain impact on splicing, mRNA analysis is helpful for ascertainment of alternative splicing and accurate interpretation of pathogenicity.<\/jats:p><\/jats:sec>","DOI":"10.1111\/ahg.12400","type":"journal-article","created":{"date-parts":[[2020,8,10]],"date-time":"2020-08-10T20:38:42Z","timestamp":1597091922000},"page":"456-462","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["RT\u2010PCR analysis of mRNA revealed the splice\u2010altering effect of rare intronic variants in monogenic disorders"],"prefix":"10.1111","volume":"84","author":[{"given":"Xia","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenjuan","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huili","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiantao","family":"Ye","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanjie","family":"Fan","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongguo","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Pediatric Endocrinology\/Genetics, Xinhua Hospital Shanghai Jiao Tong University School of Medicine Shanghai Institute for Pediatric Research  Shanghai China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,8,10]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.23626"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.23878"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2009.02.011"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.15252\/embr.201541116"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrneph.2017.83"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp215"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajmg.a.38348"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/24.17.3439"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/humu.21391"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1090100"},{"key":"e_1_2_9_12_1","article-title":"The mutational constraint spectrum quantified from variation in 141,456 humans","author":"Karczewski K. 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