Establishment of bisulfite conversion method in microcapsules for single-cell analysis.

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K14667
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: University of Yamanashi (2018); Tokyo Medical and Dental University (2016-2017)
• Principal Investigator: KOHDA Takashi 山梨大学, 大学院総合研究部, 教授 (60211893)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: メチルシトシン / エピゲノム / バイサルファイト反応 / マイクロカプセル

Outline of Final Research Achievements

The bisulfite method, which is an analytical technique for methyl cytosine, is an essential experimental method in epigenetics. However, since this reaction requires a high temperature reaction under acidic conditions, DNA cleavage occurs simultaneously with the conversion reaction. Therefore, analysis from a small amount of material was difficult. In this study, an experimental system was established in which a small amount of DNA or cells were encapsulated in alginate microcapsules, bisulfite treatment and subsequent PCR amplification were performed. This method makes it possible to handle the bisulfite treatment of trace amounts of samples much more easily and reliably, that has been previously difficult.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

哺乳類のエピゲノム制御において、DNAのシトシンメチル化はヒストン修飾とともに主要な役割を果たしている。しかしメチルシトシン解析のためのbisulfite法は微量サンプルからの解析にいくつかの困難が伴ってきた。本研究によってDNA１分子単位でbisulfite処理をマイクロカプセル中で実現しモニターすることが可能となった。この技術はbisulfite法の１細胞レベルでの解析技術開発のための重要なマイルストーンとなることが期待できる。