Strand-selective base editing of human mitochondrial DNA using mitoBEs - PKU1898
for all TALE-array recognition sequences in this manuscript). Ligated plasmids were transformed into Trans1-T1 chemically competent cells and subjected to Sanger sequencing to analyze the identity of the constructs . Final plasmids were prepared for cell transfection..
. The parameters were as follows: -t 8, -U [AG], -n 0.0, -T 6-6, -e, -d, and -u. All the significant base conversions within the targeted regions calculated by Fisher’s exact test were considered edits made by the mitoBE. The mutations that appeared in the control and experimental groups simultaneously were considered to be due to single nucleotide polymorphisms.The quality control of whole-genome sequencing was conducted using FastQC , and adapters were removed by fastp .
GM10742 cells were seeded in a polylysine-coated Seahorse XF Cell Culture Microplate using the appropriate cell culture growth medium before analysis in a Seahorse XFe24 Analyzer . Analysis was performed in Seahorse XF RPMI 1640 pH 7.4 with 10 mM glucose , 2 mM-glutamine and 1 mM sodium pyruvate . The mitochondrial function of the cells was analyzed by sequential injections of modulators .Quantitative PCR reactions were performed on a LightCycler 96 Instrument using SYBR Green .
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