Validation of the use of an artificial mitochondrial reporter DNA vector containing a Cytomegalovirus promoter for mitochondrial transgene expression

Biomaterials
Yuma YamadaHideyoshi Harashima

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 expres...Continue Reading

Citations

May 17, 2018·Mitochondrial DNA. Part A. DNA Mapping, Sequencing, and Analysis·N VerechshaginaI Mazunin
Oct 10, 2019·Nucleosides, Nucleotides & Nucleic Acids·Yuma YamadaHideyoshi Harashima
Jan 18, 2020·Biomolecules·Yuma YamadaHideyoshi Harashima
Sep 14, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yoon-Ha Jang, Kwang-Il Lim
Dec 29, 2020·Journal of Controlled Release : Official Journal of the Controlled Release Society·Yusuke SatoHideyoshi Harashima
Sep 12, 2020·Journal of Controlled Release : Official Journal of the Controlled Release Society·Yuma YamadaHideyoshi Harashima
Jun 3, 2021·Pharmaceutics·Yoon-Ha JangKwang-Il Lim
Jan 4, 2022·ACS Biomaterials Science & Engineering·Naoto Yoshinaga, Keiji Numata

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