Improvement of cDNA conversion efficiency to detect full-length sequences of extremely low amounts of RNA

2021 Fiscal Year Final Research Report

• Project/Area Number: 19H03214
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Tokyo Medical and Dental University (2021); Institute of Physical and Chemical Research (2019-2020)
• Principal Investigator: SASAGAWA YOHEI 東京医科歯科大学, 難治疾患研究所, 准教授 (10404344)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: cDNA変換 / 長鎖RNA配列

Outline of Final Research Achievements

The project's aim is to develop a highly sensitive method for detecting full-length sequences of extremely low amounts of RNA present in intracellular organelles and other samples of less than single-cell. Therefore, in this study, I attempted to improve cDNA conversion efficiency and the sequencing method to detect full-length cDNA sequence. As a result, I discovered several candidate factors that improve the efficiency of cDNA conversion. I also succeeded in establishing a workflow to efficiently analyze amplified cDNA on a long-read sequencer.

Free Research Field

ゲノミクス

Academic Significance and Societal Importance of the Research Achievements

1細胞以下の超微量RNAの検出の要望は高まっておりcDNA変換効率の改善が必要であるが、ほとんど改善されないまま使われていた。またcDNA変換は、基礎的な分子生物学的な技術であり、その改善は、微量RNAを扱う研究者・産業界など広範囲に影響を及ぼすと考えられる。本研究での知見は、幅広い分野に直接的な技術貢献をするとともに、超微量RNAからの完全長RNAを可能にし、未知RNAの発見につながると期待される。