Name your collection: Name must be less than characters. Format: Summary Summary text Abstract Abstract text. Cytidine deamination yields the mer product.
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Conflict of interest statement Competing interests The Broad Institute and the University of Washington have filed provisional patent applications on base-editing systems described in this study, listing B. Figures Extended Data Fig. Analysis of…. Analysis of the bactericidal activity of DddA and its activity…. Sequencing coverage was —fold.
Rif Rrifampicin resistant colonies. Create a file for external citation management software Create file Cancel. Genomic DNA was collected 3 days bdf or 6 days aceg post-transfection.
Untreated cells were used as controls. Send even when there aren't any new. Images are representative of 3 independent biological replicates.
Base-editing efficiencies and indel frequencies of all DddA tox splits…. Add to Collections Create a new collection Add to an existing collection.
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Editing efficiencies are measured by targeted amplicon sequencing 3 days post-transfection ab or six days post-transfection c see Methods for primer sequences. The Broad Institute and the University of Washington have filed provisional patent applications on base-editing systems described in this study, listing B. Extended Data Fig. DddA tox deaminates cytidines in bacteria with strong sequence context…. Add to My Bibliography My Bibliography. Off-target editing…. Test search terms. Two fusion orientations aureus-N or aureus-C are possible for a given split.
All cells were collected 6 days post-transfection. Optional text in :. High- or low-confidence calls indicated in parentheses were made according to the standard parameters of the prediction software. Please enable it to take advantage of the complete set of features! Display options Display options. Target cytosines are in purple. TALE arrays…. All figures Unoptimized mitoTALE—split….
The average coverage of each base was 5, to 9,fold see Supplementary Data 1. Editing efficiencies are shown on the right. Unable to load your delegates due to an error Please try again. Would you like updates of new search ? Individual P values are listed in Supplementary Table 7.
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Full text links Cite Display options Display options. As a consequence, manipulation of mtDNA to date has been limited to the targeted destruction of the mitochondrial genome by deer nucleases 9, Here we describe an interbacterial toxin, which we name DddA, that catalyses the deamination of cytidines within dsDNA.
PAM, protospacer adjacent motif. DdCBE editing….
Mitochondrial localization was followed using MitoTracker magenta. Editing efficiencies and indel frequencies for the possible combinations of UGI positions and linker lengths are shown. Bits reflect sequence conservation at a given position. DdCBE editing at five…. Abstract Bacterial toxins represent a vast reservoir of biochemical diversity that can be repurposed for biomedical applications.
Values are derived from eight independent biological replicates. DddA tox…. Stalling mtDNA…. Gels are representative of three replicates. Target cytosines and TALE-binding sites are shown in purple and blue, respectively. Stalling mtDNA replication impairs mitochondrial base editing in human cells.
Send at most: 1 item 5 items 10 items 20 items 50 items items items. Save Cancel. Target cytosines are shown in purple. DddA is a double-stranded…. Which day?
Create file Cancel. Cytidine deamination le to products P with increased mobility. Competing interests. Search: Search. Advanced Clipboard. Mitochondrial genome-wide off-target DNA…. For a — easterisks indicate ificant editing based on a comparison between indicated time points. Your saved search Name of saved search:. Frequency: Monthly Weekly Daily.
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Predicted effects…. Such proteins include a group of predicted interbacterial toxins of the deaminase superfamily, members of which have found application in gene-editing techniques 1,2. Search terms:. Choose a collection: Unable to load your collection due to an error Please try again.
: change. The experiment was repeated three times with similar. Nucleotide mismatches between the mtDNA and nuclear pseudogene are in red. We used DdCBEs to model a disease-associated mtDNA mutation in human cells, resulting in changes in respiration rates and oxidative phosphorylation. Add Cancel. Target cytosines are shown in purple and TALE binding sites are shown in blue. of items displayed: 5 10 15 20 50 Create RSS Cancel.
TALE binding sites begin at N 0 and are shown in blue. RSS Link Copy.
Luminescence values were normalized to the untreated control. The dashed line is drawn at 0. Base-editing efficiencies…. Effect of…. Edited cytosines are boxed. FCCP, carbonyl cyanide-4 trifluoromethoxy phenylhydrazone. Non-toxic split-DddA tox halves….
Bacterial toxins represent a vast reservoir of biochemical diversity that can be repurposed for biomedical applications. MeSH and other data. Other mutations are represented on the plus strand. citation Subject: 1 selected item: - PubMed. Send Cancel. Clipboard, Search History, and several other advanced features are temporarily unavailable. Mutations occurring in protein-coding regions of mtDNA were further categorized into synonymous, missense or nonsense mutations. We engineered split-DddA halves that are non-toxic and inactive until brought together on target DNA by adjacently bound programmable DNA-binding proteins.
Cells were collected 3 days post-transfection for DNA sequencing.
Deamination activity in a cleaved product P. Primer extension was performed in reactions with ddGTP to terminate primer extension at cytidines.