RRC ID 61124
Author Yamada Y, Ishikawa T, Harashima H.
Title Validation of the use of an artificial mitochondrial reporter DNA vector containing a Cytomegalovirus promoter for mitochondrial transgene expression.
Journal Biomaterials
Abstract Mitochondria have their own gene expression system that is independent of the nuclear system, and control cellular functions in cooperation with the nucleus. While a number of useful technologies for achieving nuclear transgene expression have been reported, only a few have focused on mitochondria. In this study, we validated the utility of an artificial mitochondrial DNA vector with a virus promoter on mitochondrial transgene expression. We designed and constructed pCMV-mtLuc (CGG) that contains a CMV promotor derived from Cytomegalovirus and an artificial mitochondrial genome with a NanoLuc (Nluc) luciferase gene that records adjustments to the mitochondrial codon system. Nluc luciferase activity measurements showed that the pCMV-mtLuc (CGG) efficiently produced the Nluc luciferase protein in human HeLa cells. Moreover, we optimized the mitochondrial transfection of pCMV-mtLuc (CGG) using a MITO-Porter system, a liposome-based carrier for mitochondrial delivery via membrane fusion. As a result, we found that transfection of pCMV-mtLuc (CGG) by MITO-Porter modified with the KALA peptide (cationic amphipathic cell-penetrating peptide) showed a high mitochondrial transgene expression. The developed mitochondrial transgene expression system represents a potentially useful tool for the fields of nanoscience and nanotechnology for controlling the intracellular microenvironment via the regulation of mitochondrial function and promises to open additional innovative research fields of study.
Volume 136
Pages 56-66
Published 2017-8-1
DOI 10.1016/j.biomaterials.2017.05.016
PII S0142-9612(17)30328-9
PMID 28521201
MeSH Animals Cytomegalovirus / genetics* DNA, Mitochondrial / genetics* Gene Expression Genes, Reporter Genetic Vectors / genetics* HeLa Cells Humans Luciferases / genetics* Male Mice, Inbred C57BL Mitochondria / genetics* Promoter Regions, Genetic* Transfection / methods Transgenes*
IF 10.273
Resource
Human and Animal Cells HeLa