Development of membrane-permeable oligo nucleotide and clarification of the uptake mechanism

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K14750
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: Nagoya University
• Principal Investigator: HIRAOKA Haruka 名古屋大学, 理学研究科, 特任助教 (70880053)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 膜透過性核酸 / ジスルフィド修飾 / 核酸医薬 / 筋ジストロフィー

Outline of Final Research Achievements

In this study, we clarified the timescale of membrane permeabilization of MPON, Membrane Permeable OligoNucleotides modified with disulfide units, and identified the candidate responsible protein that specifically binds to MPON and promotes the cell transduction by mass spectrometry. Furthermore, we found that MPON is translocated into the nucleus over time after cell transduction, which led to the idea that MPON could be applied to the treatment of muscular dystrophy caused by splicing abnormalities in the nucleus. Actually, MPON functioned in alteration of splicing pattern in a mouse model of the disease. We also developed improved next-generation MPONs with different modification groups and evaluated their functions.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

研究室で開発した細胞膜透過性核酸MPONが細胞核内へと移行することが分かり、核内で起こる選択的スプライシングのパターンを高い効率で変化させることがマウスで確かめられたことから、筋ジストロフィーの原因となるスプライシング異常を是正し疾患治療に適用できる可能性が示された。また、MPONの細胞膜透過過程および細胞内動態の観察に基づき利点とさらなる改善点を明確にしたことで、改良型の次世代MPONの開発を大きく進展させた。核酸医薬としての医学的応用は勿論、膜透過による核酸材料の効率的な運搬と供給を可能にするMPONは人工細胞における核酸・タンパク質の機能解析への適用も期待され、学術的意義も高い。