PMID- 31981894
OWN - NLM
STAT- MEDLINE
DCOM- 20210416
LR  - 20231113
IS  - 2213-2317 (Electronic)
IS  - 2213-2317 (Linking)
VI  - 30
DP  - 2020 Feb
TI  - Codon optimization is an essential parameter for the efficient allotopic 
      expression of mtDNA genes.
PG  - 101429
LID - S2213-2317(19)31063-8 [pii]
LID - 10.1016/j.redox.2020.101429 [doi]
LID - 101429
AB  - Mutations in mitochondrial DNA can be inherited or occur de novo leading to 
      several debilitating myopathies with no curative option and few or no effective 
      treatments. Allotopic expression of recoded mitochondrial genes from the nucleus 
      has potential as a gene therapy strategy for such conditions, however progress in 
      this field has been hampered by technical challenges. Here we employed codon 
      optimization as a tool to re-engineer the protein-coding genes of the human 
      mitochondrial genome for robust, efficient expression from the nucleus. All 13 
      codon-optimized constructs exhibited substantially higher protein expression than 
      minimally-recoded genes when expressed transiently, and steady-state mRNA levels 
      for optimized gene constructs were 5-180 fold enriched over recoded versions in 
      stably-selected wildtype cells. Eight of thirteen mitochondria-encoded oxidative 
      phosphorylation (OxPhos) proteins maintained protein expression following stable 
      selection, with mitochondrial localization of expression products. We also 
      assessed the utility of this strategy in rescuing mitochondrial disease cell 
      models and found the rescue capacity of allotopic expression constructs to be 
      gene specific. Allotopic expression of codon optimized ATP8 in disease models 
      could restore protein levels and respiratory function, however, rescue of the 
      pathogenic phenotype for another gene, ND1 was only partially successful. These 
      results imply that though codon-optimization alone is not sufficient for 
      functional allotopic expression of most mitochondrial genes, it is an essential 
      consideration in their design.
CI  - Copyright (c) 2020 The Authors. Published by Elsevier B.V. All rights reserved.
FAU - Lewis, Caitlin J
AU  - Lewis CJ
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA.
FAU - Dixit, Bhavna
AU  - Dixit B
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA.
FAU - Batiuk, Elizabeth
AU  - Batiuk E
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA.
FAU - Hall, Carter J
AU  - Hall CJ
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA.
FAU - O'Connor, Matthew S
AU  - O'Connor MS
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA. Electronic address: matthew.oconnor@sens.org.
FAU - Boominathan, Amutha
AU  - Boominathan A
AD  - Department of Mitochondrial Research, SENS Research Foundation, Mountain View, 
      CA, 94041, USA. Electronic address: amutha.boominathan@sens.org.
LA  - eng
PT  - Journal Article
DEP - 20200111
PL  - Netherlands
TA  - Redox Biol
JT  - Redox biology
JID - 101605639
RN  - 0 (DNA, Mitochondrial)
RN  - 0 (Mitochondrial Proteins)
RN  - EC 1.6.99.3 (NADH Dehydrogenase)
RN  - EC 1.6.99.3 (NADH dehydrogenase subunit 1, human)
RN  - EC 3.6.3.- (Mitochondrial Proton-Translocating ATPases)
SB  - IM
MH  - Animals
MH  - Codon Usage
MH  - Computational Biology/*methods
MH  - DNA, Mitochondrial/genetics
MH  - Gene Expression
MH  - HEK293 Cells
MH  - Humans
MH  - Mice
MH  - Mitochondria/*genetics
MH  - Mitochondrial Diseases/*genetics
MH  - Mitochondrial Proteins/genetics
MH  - Mitochondrial Proton-Translocating ATPases/*genetics
MH  - Models, Biological
MH  - *Mutation
MH  - NADH Dehydrogenase/*genetics
PMC - PMC6976934
OTO - NOTNLM
OT  - ATP synthase
OT  - Allotopic expression
OT  - Codon optimization
OT  - Gene therapy
OT  - Gene transfer
OT  - Mitochondrial DNA (mtDNA)
OT  - Mitochondrial disease
OT  - Mitochondrial respiratory chain complex
OT  - Protein expression
COIS- Declaration of competing interest The authors declare no competing interests.
EDAT- 2020/01/26 06:00
MHDA- 2021/04/17 06:00
PMCR- 2020/01/11
CRDT- 2020/01/26 06:00
PHST- 2019/09/07 00:00 [received]
PHST- 2019/12/29 00:00 [revised]
PHST- 2020/01/10 00:00 [accepted]
PHST- 2020/01/26 06:00 [pubmed]
PHST- 2021/04/17 06:00 [medline]
PHST- 2020/01/26 06:00 [entrez]
PHST- 2020/01/11 00:00 [pmc-release]
AID - S2213-2317(19)31063-8 [pii]
AID - 101429 [pii]
AID - 10.1016/j.redox.2020.101429 [doi]
PST - ppublish
SO  - Redox Biol. 2020 Feb;30:101429. doi: 10.1016/j.redox.2020.101429. Epub 2020 Jan 
      11.