20225-06-20 北海道大学,科学技術振興機構
MITO-Porter を用いたミトコンドリアへのゲノム編集装置(RNP)送達の概念図
<関連情報>
- https://www.hokudai.ac.jp/news/2025/06/post-1929.html
- https://www.hokudai.ac.jp/news/pdf/250620_pr.pdf
- https://www.nature.com/articles/s41598-025-03671-8
ミトコンドリアDNA(mt-Atp8)にm.7778G>T変異を有する細胞のミトコンドリアへCRISPR/Cas9システムを直接送達する革新的な脂質ナノカプセル Lipid nanoparticle delivery of the CRISPR/Cas9 system directly into the mitochondria of cells carrying m.7778G>T mutation in MtDNA (mt-Atp8)
Kaede Norota,Sen Ishizuka,Misa Hirose,Yusuke Sato,Masatoshi Maeki,Manabu Tokeshi,Saleh M. Ibrahim,Hideyoshi Harashima & Yuma Yamada
Scientific Reports Published:19 June 2025
DOI:https://doi.org/10.1038/s41598-025-03671-8
Abstract
Mitochondrial genome mutations are associated with various diseases and gene therapy targeted to mitochondria has the potential to effectively treat such diseases. Here, we targeted a point mutation in mitochondrial DNA (mtDNA) that can cause mitochondrial diseases via delivery of the clustered, regularly interspaced, short palindromic repeats/Cas9 (CRISPR/Cas9) system to mitochondria using an innovative lipid nanoparticle (LNP) delivery system. To overcome the major barrier of the mitochondrial membrane structure, we investigated a strategy to deliver ribonucleoprotein (RNP) directly to mitochondria via membrane fusion using MITO-Porter, a mitochondria-targeting lipid nanoparticle. First, we constructed RNP-MITO-Porter, in which an RNP was loaded into MITO-Porter using a microfluidic device. Sequence-specific double-strand breaks were confirmed when the constructed RNP-MITO-Porter was applied to isolated mitochondria. Next, the RNP-MITO-Porter was applied to HeLa cells, and a portion of the RNP-MITO-Porter was colocalized with mitochondria and caused sequence-specific double-strand breaks in mtDNA. Finally, RNP-MITO-Porter was successfully delivered to mitochondria of cells derived from a mouse carrying a point mutation (m.7778G > T) in mtDNA (mt-Atp8) (LMSF-N-MTFVB cells), and created double-strand breaks at the target sequence. RNP-MITO-Porter is expected to contribute significantly to the clinical application of mitochondrion-targeted gene therapy.
