GSE173689
Detect-seq on transfected HEK293T cells [ATAC-seq data and in situ ChIP-seq data]
We used a new technology, named Detect-seq, to perform genome sequencing on transfected HEK293T cells to see DdCBEs' off-target mutations. Besides, targeted-amplicon sequencing, ATAC-seq and in situ ChIP-seq data were applied to validate the results of Detect-seq. Overall design: We applied DdCBEs to generate on-target editings on mt-DNA to see DdCBEs' editing performance and evaluate their effects on the whole mitochondrial genome. These are the ATAC-seq part and in situ ChIP-seq part . ATAC-seq was performed as previously reported. 10,000 viable cells were pelleted, lysed and the genomic DNA was tagmented with 2.5µl self-assembled Tn5 transposase. The tagmented DNA was purified using the DNA Clean and Concentrator-5 Kit (Vistech) and eluted in 20µl ultrapure water. All 20µl eluate was amplified by 12 cycles in a total volume of 50µl and the final library was purified and size-selected using Agencourt AMPure XP beads (Beckman Coulter). Low-input in situ ChIP was performed as previously described with minor modifications mentioned in our paper.