| RRC ID |
55981
|
| 著者 |
Ishikawa T, Somiya K, Munechika R, Harashima H, Yamada Y.
|
| タイトル |
Mitochondrial transgene expression via an artificial mitochondrial DNA vector in cells from a patient with a mitochondrial disease.
|
| ジャーナル |
J Control Release
|
| Abstract |
To achieve mitochondrial gene therapy, developing a mitochondrial transgene expression system that produces therapeutic proteins in mitochondria of disease cells is essential. We previously reported on the design of pCMV-mtLuc (CGG) containing a CMV promotor and a NanoLuc (Nluc) luciferase gene that records adjustments to the mitochondrial codon system, and showed that the mitochondrial transfection of pCMV-mtLuc (CGG) resulted in the efficient production of the Nluc luciferase protein in human HeLa cells. This mitochondrial transfection was achieved using a MITO-Porter, a liposome-based carrier for delivering a cargo to mitochondria via membrane fusion. We report herein that mitochondrial transfection using the MITO-Porter results in mitochondrial transgene expression in G625A fibroblasts obtained from a patient with a mitochondrial disease. We investigated the effect of promoters and the basic structure of pCMV-mtLuc (CGG) on gene expression efficiency, and were able to construct a high performance mitochondrial DNA vector, pCMV-mtLuc (CGG) [hND4] that contains a human mitochondrial endogenous gene. We also constructed an RP/KALA-MITO-Porter composed of the KALA peptide (cell-penetrating peptide) with a mitochondrial RNA aptamer to enhance cellular uptake and mitochondrial targeting. Finally, the mitochondrial transfection of pCMV-mtLuc (CGG) [hND4] in G625A fibroblasts using the RP/KALA-MITO-Porter resulted in strong mitochondrial transgene expression.
|
| 巻・号 |
274
|
| ページ |
109-117
|
| 公開日 |
2018-3-28
|
| DOI |
10.1016/j.jconrel.2018.02.005
|
| PII |
S0168-3659(18)30065-8
|
| PMID |
29408532
|
| MeSH |
Animals
DNA, Mitochondrial / administration & dosage*
DNA, Mitochondrial / genetics
Gene Transfer Techniques*
Genetic Therapy / methods
Humans
Male
Mice, Inbred C57BL
Mitochondria / genetics*
Mitochondria / metabolism
Mitochondrial Diseases / therapy*
Transfection
Transgenes
|
| IF |
7.901
|
| 引用数 |
15
|
| リソース情報 |
| ヒト・動物細胞 |
HeLa |
