Abstract：
:Precise editing is essential for biomedical research and gene therapy. Yet, homology-directed genome modification is limited by the requirements for genomic lesions, homology donors and the endogenous DNA repair machinery. Here we engineered programmable cytidine deaminases and test if we could introduce site-specific cytidine to thymidine transitions in the absence of targeted genomic lesions. Our programmable deaminases effectively convert specific cytidines to thymidines with 13% efficiency in Escherichia coli and 2.5% in human cells. However, off-target deaminations were detected more than 150 bp away from the target site. Moreover, whole genome sequencing revealed that edited bacterial cells did not harbour chromosomal abnormalities but demonstrated elevated global cytidine deamination at deaminase intrinsic binding sites. Therefore programmable deaminases represent a promising genome editing tool in prokaryotes and eukaryotes. Future engineering is required to overcome the processivity and the intrinsic DNA binding affinity of deaminases for safer therapeutic applications.
journal_name
Nat Communjournal_title
Nature communicationsauthors
Yang L,Briggs AW,Chew WL,Mali P,Guell M,Aach J,Goodman DB,Cox D,Kan Y,Lesha E,Soundararajan V,Zhang F,Church Gdoi
10.1038/ncomms13330subject
Has Abstractpub_date
2016-11-02 00:00:00pages
13330issn
2041-1723pii
ncomms13330journal_volume
7pub_type
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