Chapter title |
Delivery of mtZFNs into Early Mouse Embryos
|
---|---|
Chapter number | 16 |
Book title |
Zinc Finger Proteins
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8799-3_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8798-6, 978-1-4939-8799-3
|
Authors |
Beverly J. McCann, Andy Cox, Payam A. Gammage, James B. Stewart, Magdalena Zernicka-Goetz, Michal Minczuk, McCann, Beverly J., Cox, Andy, Gammage, Payam A., Stewart, James B., Zernicka-Goetz, Magdalena, Minczuk, Michal |
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. |
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