Development of metal-responsive DNA supramolecules based on enzymatic DNA synthesis

2021 Fiscal Year Final Research Report

• Project/Area Number: 18H02081
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: Takezawa Yusuke 東京大学, 大学院理学系研究科(理学部), 助教 (70508598)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: DNA / 人工DNA / DNAポリメラーゼ / DNAリガーゼ / DNAzyme / DNAナノテクノロジー / 金属錯体型塩基対 / 超分子化学

Outline of Final Research Achievements

We have developed facile enzymatic methods to synthesize DNA strands containing a ligand-type hydroxypyridone (H) nucleotide, which forms a Cu(II)-mediated base pair (H-Cu(II)-H). The H-modified oligonucleotides were synthesized by a primer extension reaction using two DNA polymerases or by a two-step reaction with polymerases and ligases. The enzymatic synthesis was subsequently applied to the development of metal-responsive allosteric DNAzymes, whose catalytic activity can be regulated by the formation of metal-mediated base pairs. The rational design strategy and the easy enzymatic synthesis established in this study provide a versatile way to develop a variety of metal-responsive DNA materials based on metal-mediated artificial base pairing.

Free Research Field

超分子化学

Academic Significance and Societal Importance of the Research Achievements

本研究では、DNA合成酵素を用いた金属配位子型人工DNAの酵素合成法の開発と、それに基づく金属イオン応答性DNAzymeの開発を行った。金属応答性DNA鎖の簡便な合成が可能になった点、および既存の機能性核酸を基に配列を合理設計できる点が特長であり、他の機能性核酸やDNAナノ構造体の構造・機能制御へも応用できる成果である。特に一対の非天然塩基対の導入のみで活性制御が可能となり、最小限の修飾・改変による核酸の機能化という点で大きな意義がある。生体微量金属を含む種々の金属イオンに応答する機能性核酸も創出できると考えられ、DNAナノテクノロジーや生命金属科学の基礎分子技術としての波及効果も期待される。