{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T20:51:41Z","timestamp":1740171101418,"version":"3.37.3"},"reference-count":26,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2017,2,8]],"date-time":"2017-02-08T00:00:00Z","timestamp":1486512000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science and Technology","doi-asserted-by":"publisher","award":["16K09962"],"award-info":[{"award-number":["16K09962"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003478","name":"Ministry of Health, Labour and Welfare","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003478","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009619","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":[[2017,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>\u03b2\u2010ketothiolase (T2, gene symbol <jats:italic><jats:styled-content style=\"fixed-case\">ACAT<\/jats:styled-content>1<\/jats:italic>) deficiency is an autosomal recessive disorder, affecting isoleucine and ketone body metabolism. We encountered a patient (<jats:styled-content style=\"fixed-case\">GK<\/jats:styled-content>03) with T2 deficiency whose T2 <jats:styled-content style=\"fixed-case\">mRNA<\/jats:styled-content> level was &lt;10% of the control, but in whom a previous routine <jats:styled-content style=\"fixed-case\">cDNA<\/jats:styled-content> analysis had failed to find any mutations. Genomic <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>\u2010direct sequencing showed homozygosity for c.941\u20109T&gt;A in the polypyrimidine stretch at the splice acceptor site of intron 9 of <jats:italic><jats:styled-content style=\"fixed-case\">ACAT<\/jats:styled-content>1<\/jats:italic>. Initially, we regarded this variant as not being disease\u2010causing by a method of predicting the effect of splicing using in silico tools. However, based on other findings of exon 10 splicing, we eventually hypothesized that this mutation causes exon 10 skipping.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">cDNA<\/jats:styled-content> analysis was performed using <jats:styled-content style=\"fixed-case\">GK<\/jats:styled-content>03's fibroblasts treated with\/without cycloheximide (<jats:styled-content style=\"fixed-case\">CHX<\/jats:styled-content>), since exon 10 skipping caused a frameshift and nonsense\u2010mediated <jats:styled-content style=\"fixed-case\">mRNA<\/jats:styled-content> decay (<jats:styled-content style=\"fixed-case\">NMD<\/jats:styled-content>). Minigene splicing experiment was done to confirm aberrant splicing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">cDNA<\/jats:styled-content> analysis using fibroblasts cultured with cycloheximide indeed showed the occurrence of exon 10 skipping. A minigene splicing experiment clearly showed that the c.941\u20109T&gt;A mutant resulted in transcripts with exon 10 skipping. There are few reports describing that single\u2010nucleotide substitutions in polypyrimidine stretches of splice acceptor sites cause aberrant splicing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We showed that c.941\u20109T&gt;A induces aberrant splicing in the <jats:italic><jats:styled-content style=\"fixed-case\">ACAT<\/jats:styled-content>1<\/jats:italic> gene. Our ability to predict the effects of mutations on splicing using in silico tools is still limited. <jats:styled-content style=\"fixed-case\">cDNA<\/jats:styled-content> analysis and minigene splicing experiments remain useful alternatives to reveal splice defects.<\/jats:p><\/jats:sec>","DOI":"10.1002\/mgg3.275","type":"journal-article","created":{"date-parts":[[2017,2,8]],"date-time":"2017-02-08T05:15:15Z","timestamp":1486530915000},"page":"177-184","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Single\u2010nucleotide substitution T to A in the polypyrimidine stretch at the splice acceptor site of intron 9 causes exon 10 skipping in the <i><scp>ACAT<\/scp>1<\/i> gene"],"prefix":"10.1002","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7704-6259","authenticated-orcid":false,"given":"Hideo","family":"Sasai","sequence":"first","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuka","family":"Aoyama","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"},{"name":"Department of Biomedical Sciences College of Life and Health Sciences Chubu University Kasugai Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroki","family":"Otsuka","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elsayed","family":"Abdelkreem","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"},{"name":"Department of Pediatrics Faculty of Medicine Sohag University Sohag Egypt"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mina","family":"Nakama","sequence":"additional","affiliation":[{"name":"Division of Clinical Genetics Gifu University Hospital Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomohiro","family":"Hori","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hidenori","family":"Ohnishi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lesley","family":"Turner","sequence":"additional","affiliation":[{"name":"Discipline of Genetics Memorial University of Newfoundland St John's NF Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshiyuki","family":"Fukao","sequence":"additional","affiliation":[{"name":"Department of Pediatrics Graduate School of Medicine Gifu University Gifu Japan"},{"name":"Division of Clinical Genetics Gifu University Hospital Gifu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,2,8]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-013-2500-z"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/mus.21937"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(71)90605-2"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp215"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI114946"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1203\/00006450-199606000-00020"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1203\/00006450-199710000-00013"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-1004(1998)12:4<245::AID-HUMU5>3.0.CO;2-E"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2008.04.014"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2010.03.012"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2013.07.004"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10545-014-9704-9"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.10-6933"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(91)90623-J"},{"key":"e_1_2_6_16_1","first-page":"2675","article-title":"The missing puzzle piece: splicing mutations","volume":"6","author":"Lewandowska M. 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