Delivery of mtZFNs into early mouse embryos
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Authors
McCann, Beverly
Cox, Andy
Gammage, Payam
Stewart, James
Journal Title
Zinc Finger Proteins
ISBN
978-1-4939-8798-6
Publisher
Humana Press
Volume
1867
Pages
215-228
Language
English
Type
Book chapter
This Version
NA
Metadata
Show full item recordCitation
McCann, B., Cox, A., Gammage, P., Stewart, J., Zernicka-Goetz, M., & Minczuk, M. Delivery of mtZFNs into early mouse embryos. Humana Press, Zinc Finger Proteins. [Book chapter]. https://doi.org/10.1007/978-1-4939-8799-3_16
Abstract
Mitochondrial diseases often result from mutations in the mitochondrial genome (mtDNA). In most cases, mutant mtDNA coexists with wild-type mtDNA, resulting in heteroplasmy. One potential future approach to treat heteroplasmic mtDNA diseases is the specific elimination of pathogenic mtDNA mutations, lowering the level of mutant mtDNA below pathogenic thresholds. Mitochondrially-targeted zinc finger nucleases (mtZFNs) have been demonstrated to specifically target and introduce double-strand breaks in mutant mtDNA, facilitating substantial shifts in heteroplasmy. One application of mtZFN technology, in the context of heteroplasmic mtDNA disease, is delivery into the heteroplasmic oocyte or early embryo to eliminate mutant mtDNA, preventing transmission of mitochondrial diseases through the germline. Here we describe a protocol for efficient production of mtZFN mRNA in vitro, and delivery of these into 0.5 dpc mouse embryos to elicit shifts of mtDNA heteroplasmy.
Sponsorship
Champ Foundation (unknown)
MRC (MC_U105697135)
MRC (MC_UU_00015/4)
Embargo Lift Date
2100-01-01
Identifiers
External link: https://link.springer.com/protocol/10.1007%2F978-1-4939-8799-3_16#copyrightInformation
External DOI: https://doi.org/10.1007/978-1-4939-8799-3_16
This record's DOI: https://doi.org/10.17863/CAM.37918
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Licence:
http://www.rioxx.net/licenses/all-rights-reserved