Elucidation of a mechanism of tissue-specific alternative splicing by using analysis of splicing order.

2015 Fiscal Year Final Research Report

• Project/Area Number: 26860793
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Pediatrics
• Research Institution: Gifu University
• Principal Investigator: HORI Tomohiro 岐阜大学, 医学部附属病院, 助教 (90456525)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: スプライシングオーダー / 選択的スプライシング / 遺伝学

Outline of Final Research Achievements

Our strategy was to use RT-PCR of hnRNA to detect the presence or absence of spliced exon clusters in RNA intermediates (SECRIs) comprising sequential exons. The splice donor site mutation c.1248+5g>a (IVS13) of the OXCT1 gene resulted predominantly in skipping of exons 12 and 13 in skin fibroblasts from a patient with succinyl-CoA:3-ketoacid CoA transferase (SCOT) deficiency. We compared hnRNA intermediates between controls' and the patient's fibroblasts derived from skin, kidney and liver. In controls, intron 11 was the last intron to be spliced and the removal of intron 12 was also rather slow and occurred after the removal of intron 13 in a major pathway. The mutation in the patient’s cells resulted in retention of intron 13, thus causing the retention of introns 12 and 11. Now, we have been conducting studies about splicing order in fibroblasts derived from hepatocyte and the gene responsible for mitochondrial acetoacetyl-CoA thiolase deficiency.

Free Research Field

小児科学