Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (29)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Srivastava, S.
Right arrow Articles by Moraes, C. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Srivastava, S.
Right arrow Articles by Moraes, C. T.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Human Molecular Genetics, 2001, Vol. 10, No. 26 3093-3099
© 2001 Oxford University Press

Manipulating mitochondrial DNA heteroplasmy by a mitochondrially targeted restriction endonuclease

Sarika Srivastava1 and Carlos T. Moraes1,2,+

1Department of Cell Biology and Anatomy and 2Department of Neurology, University of Miami School of Medicine, 1095 NW 14th Terrace, Miami, FL 33136, USA

Mutations in the mitochondrial DNA (mtDNA) can cause a variety of human diseases. In most cases, such mutations are heteroplasmic (i.e. mutated and wild-type mtDNA coexist) and a small percentage of wild-type sequences can have a strong protective effect against a metabolic defect. Because a genetic approach to correct mtDNA mutations is not currently available, the ability to modulate heteroplasmy would have a major impact in the phenotype of many patients with mitochondrial disorders. We show here that a restriction endonuclease targeted to mitochondria has this ability. A mitochondrially targeted PstI degraded mtDNA harboring PstI sites, in some cases leading to a complete loss of mitochondrial genomes. Recombination between DNA ends released by PstI was not observed. When expressed in a heteroplasmic rodent cell line, containing one mtDNA haplotype with two sites for PstI and another haplotype having none, the mitochondrial PstI caused a significant shift in heteroplasmy, with an accumulation of the mtDNA haplotype lacking PstI sites. These experiments provide proof of the principle that restriction endonucleases are feasible tools for genetic therapy of a sub-group of mitochondrial disorders. Although this approach is limited by the presence of mutation-specific restriction sites, patients with neuropathy, ataxia and retinitis pigmentosa (NARP) could benefit from it, as the T8399G mutation creates a unique restriction site that is not present in wild-type human mitochondrial DNA.

+ To whom correspondence should be addressed. Tel: +1 305 243 5858; Fax: +1 305 243 3914; Email: cmoraes@med.miami.edu


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Hum Mol GenetHome page
A. Malena, E. Loro, M. Di Re, I. J. Holt, and L. Vergani
Inhibition of mitochondrial fission favours mutant over wild-type mitochondrial DNA
Hum. Mol. Genet., September 15, 2009; 18(18): 3407 - 3416.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
H. Xu, S. Z. DeLuca, and P. H. O'Farrell
Manipulating the Metazoan Mitochondrial Genome with Targeted Restriction Enzymes
Science, July 25, 2008; 321(5888): 575 - 577.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Minczuk, M. A. Papworth, J. C. Miller, M. P. Murphy, and A. Klug
Development of a single-chain, quasi-dimeric zinc-finger nuclease for the selective degradation of mutated human mitochondrial DNA
Nucleic Acids Res., July 1, 2008; 36(12): 3926 - 3938.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Kukat, C. Kukat, J. Brocher, I. Schafer, G. Krohne, I. A. Trounce, G. Villani, and P. Seibel
Generation of {rho}0 cells utilizing a mitochondrially targeted restriction endonuclease and comparative analyses
Nucleic Acids Res., April 1, 2008; 36(7): e44 - e44.
[Abstract] [Full Text] [PDF]

Home page
 Hum Mol GenetHome page
H. Sembongi, M. Di Re, M. Bokori-Brown, and I. J. Holt
The yeast Holliday junction resolvase, CCE1, can restore wild-type mitochondrial DNA to human cells carrying rearranged mitochondrial DNA
Hum. Mol. Genet., October 1, 2007; 16(19): 2306 - 2314.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
S. M. Khan, R. M. Smigrodzki, and R. H. Swerdlow
Cell and animal models of mtDNA biology: progress and prospects
Am J Physiol Cell Physiol, February 1, 2007; 292(2): C658 - C669.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. P. Bayona-Bafaluy, B. Blits, B. J. Battersby, E. A. Shoubridge, and C. T. Moraes
Rapid directional shift of mitochondrial DNA heteroplasmy in animal tissues by a mitochondrially targeted restriction endonuclease
PNAS, October 4, 2005; 102(40): 14392 - 14397.
[Abstract] [Full Text] [PDF]

Home page
 Hum Mol GenetHome page
S. Srivastava and C. T. Moraes
Double-strand breaks of mouse muscle mtDNA promote large deletions similar to multiple mtDNA deletions in humans
Hum. Mol. Genet., April 1, 2005; 14(7): 893 - 902.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.