1分子リアルタイムシークエンシング技術（PacBio）によるスプライシング欠陥の同定と特性評価

Identification and Characterization of Splicing Defects by Single-Molecule Real-Time Sequencing Technology (PacBio).

• Marco Savarese
• Talha Qureshi
• Annalaura Torella
• Pia Laine
• Teresa Giugliano
• Per Harald Jonson
• Mridul Johari
• Lars Paulin
• Giulio Piluso
• Petri Auvinen
• Vincenzo Nigro
• Bjarne Udd
• Peter Hackman

抄録

逆転写ポリメラーゼ連鎖反応(RT-PCR)は、臨床表現型や利用可能なDNA検査の結果から目的の遺伝子の診断が可能な場合や、単一変異体のスプライシング効果を評価する必要がある場合には、有用な選択肢となります。適切に最適化された後、この手順を診断の場で使用することができ、cDNA解析のワークフローを簡素化することができます。

Although DNA-sequencing is the most effective procedure to achieve a molecular diagnosis in genetic diseases, complementary RNA analyses are often required.Reverse-Transcription polymerase chain reaction (RT-PCR) is still a valuable option when the clinical phenotype and/or available DNA-test results address the diagnosis toward a gene of interest or when the splicing effect of a single variant needs to be assessed.We use Single-Molecule Real-Time sequencing to detect and characterize splicing defects and single nucleotide variants in well-known disease genes (DMD, NF1, TTN). After proper optimization, the procedure could be used in the diagnostic setting, simplifying the workflow of cDNA analysis.