Precision mitochondrial DNA editing with high-fidelity DddA-derived base editors
). We recommend the use of the T1391A variant in 1397N or 1333C when high specificity is desired over high activity, and the use of the K1389A variant in 1397N or 1333C when high activity is desired over high specificity. Note that-specific, 1397-split or 1333-split HiFi-DdCBEs with K1389A were as active as wild-type DdCBEs targeted to the same sites but were much more specific, showing a 22-fold or 21-fold difference, respectively, in average mtDNA-wide off-target editing frequencies .
Cells were not tested for mycoplasma contamination. For lipofection, cells were plated in 48-well cell culture plates at a density of 5 × 10cells per well, 18–24 h before transfection. Lipofection using Lipofectamine 2000 was performed with 500 ng of each TALE half monomer plasmid to make up 1,000 ng of total plasmid DNA. Cells were collected on day four post transfection.
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