Method for identifying positions of proteins bound to a single DNA molecule

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K14511
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 28050:Nano/micro-systems-related
• Research Institution: Nagoya University
• Principal Investigator: Naoki Azuma 名古屋大学, 工学研究科, 助教 (50823283)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Keywords: DNA一分子 / タンパク質結合位置特定 / 微小流路 / 蛍光分子局在化法 / 圧力泳動

Outline of Final Research Achievements

The purpose of this research was to realize a method for identifying the positions of proteins bound to a single DNA molecule by directly observing method. We have developed two elemental technologies. One is the stretching and immobilizing a single DNA molecule using a pressure flow in a microchannel. The other is identifying the positions of proteins bound to a single DNA molecule with 10 nm accuracy using the super-localization method, which is one of the super-resolution techiniques. The feasibility of the proposed method for identifying positions of proteins bound to a single DNAA molecule was demonstrated through experimental validation of stretching and immobilizing method and super-localization method.

Free Research Field

ナノマイクロシステム，ナノ計測，バイオ計測

Academic Significance and Societal Importance of the Research Achievements

本研究は，これまでに実現されていなかったDNA一分子の伸長・固定および光学的手法による10 nm精度の結合位置特定を実現できる独創的な手法であり，挑戦的である．従来のゲル法のように，検出に多数の分子を必要とせず，高速かつ高感度にタンパク質の結合位置特定を実現でき，DNA編集法であるCRISPR/Cas9の評価手法として貢献できる可能性があるため，社会的意義が大きい．さらに，DNA一分子で10 nm精度の分析を可能とするため，これまでの分析法に代わるDNA一分子分析法として長さ分析や塩基配列特定に展開でき，生命科学や創薬科学への波及効果も期待できる．