RRC ID 55981
Author Ishikawa T, Somiya K, Munechika R, Harashima H, Yamada Y.
Title Mitochondrial transgene expression via an artificial mitochondrial DNA vector in cells from a patient with a mitochondrial disease.
Journal 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.
Volume 274
Pages 109-117
Published 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
Times Cited 15
Human and Animal Cells HeLa