Cationized protein transduction route for mammalian cells

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K04801
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Okayama University of Science
• Principal Investigator: Futami Midori (北添翠) 岡山理科大学, 生命科学部, 准教授 (10467748)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: タンパク質細胞導入法

Outline of Final Research Achievements

By protein cationization through chemical modification, it is possible to electrostatically adsorb them onto the negatively charged cell surface and introduce arbitrary proteins into the cell. In this study, we investigated the pathway by which cationized proteins are introduced into cells.
Using the artificial transcription factor BDAD as a model protein, cationized BDAD proteins were introduced into cells treated with various endocytosis inhibitors. It was demonstrated that only cells treated with the clathrin endocytosis inhibitor showed a significant decrease in the transduction efficiency of BDAD, indicating that clathrin-mediated endocytosis is important for the introduction pathway. Furthermore, the involvement of the cell membrane protein Syndecan-4 in the cationized protein introduction was confirmed.

Free Research Field

タンパク質工学

Academic Significance and Societal Importance of the Research Achievements

カチオン化によるタンパク質導入技術ではタンパク質の種類によっては導入効果が明確に見られないものがあり、その原因究明が課題であった。今回の研究を経て、カチオン化タンパク質の導入経路が明確になり、クラスリン依存性エンドサイトーシスが優位に働いていない細胞や培養条件ではタンパク質導入が著しく難しくなることが分かった。カチオン化によるタンパク質導入法の適用範囲が明確になったことは技術を利用する場合に成否を見通す上で非常に重要であり、さらに今後は導入が難しい細胞・培養条件においても導入が可能になるような工夫を検討するうえで道筋を示す情報である。