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How gene editing can address mitochondrial DNA?

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Antonio Carusillo
Antonio Carusillo Aug 27, 2020

[1]Sun N, Youle RJ, Finkel T. The Mitochondrial Basis of Aging. Mol Cell. 2016;61(5):654-666. doi:10.1016/j.molcel.2016.01.028

[2]Heidi Chial, Ph.D. (Write Science Right) & Joanna Craig, Ph.D. (Write Science Right) © 2008 Nature Education Citation: Chial, H. & Craig, J. (2008) mtDNA and mitochondrial diseases. Nature Education 1(1):217

[3]A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity BY MARTIN JINEK, KRZYSZTOF CHYLINSKI, INES FONFARA, MICHAEL HAUER, JENNIFER A. DOUDNA, EMMANUELLE CHARPENTIER SCIENCE17 AUG 2012 : 816-821

[4]Hirakawa MP, Krishnakumar R, Timlin JA, Carney JP, Butler KS. Gene editing and CRISPR in the clinic: current and future perspectives. Biosci Rep. 2020;40(4):BSR20200127. doi:10.1042/BSR20200127

[5]Gammage PA, Moraes CT, Minczuk M. Mitochondrial Genome Engineering: The Revolution May Not Be CRISPR-Ized. Trends Genet. 2018;34(2):101-110. doi:10.1016/j.tig.2017.11.001

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Using TALEs and Base editing to edit mtDNA

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Antonio Carusillo
Antonio Carusillo Aug 28, 2020

Delivering the gRNA expression cassette as a self-replicating plasmid

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Manel Lladó Santaeularia
Manel Lladó Santaeularia Dec 06, 2020

[1]Bonnefoy N, Fox TD. Directed alteration of Saccharomyces cerevisiae mitochondrial DNA by biolistic transformation and homologous recombination. Methods Mol Biol. 2007;372:153-166. doi:10.1007/978-1-59745-365-3_11

[2]Yoo BC, Yadav NS, Orozco EM Jr, Sakai H. Cas9/gRNA-mediated genome editing of yeast mitochondria and Chlamydomonas chloroplasts. PeerJ. 2020;8:e8362. Published 2020 Jan 6. doi:10.7717/peerj.8362

[3]Yamada Y, Akita H, Kamiya H, et al. MITO-Porter: A liposome-based carrier system for delivery of macromolecules into mitochondria via membrane fusion. Biochim Biophys Acta. 2008;1778(2):423-432. doi:10.1016/j.bbamem.2007.11.002

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Antonio Carusillo
Antonio Carusillo3 months ago
Hi Manel I really like the idea. Probably In cell lines it will be possible, not quite sure in primary cells as they do not really well tolerate plasmid DNA, maybe mini-circles may come handy to this regard. Also would be interesting to see how it works for the Cas9 as protein while the gRNA is a plasmid. I do not know if there are any studies where they tried to do it. Cause I imagine the Cas9 protein being slowly degradated while the gRNA plasmid is still being transcribed. But if you can make it this will also open a new avenue about mtDNA off-target assays!

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