RT Journal Article
SR Electronic
T1 Genome Editing With Targeted Deaminases
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 066597
DO 10.1101/066597
A1 Luhan Yang
A1 Adrian W. Briggs
A1 Wei Leong Chew
A1 Prashant Mali
A1 Marc Guell
A1 John Aach
A1 Daniel Bryan Goodman
A1 David Cox
A1 Yinan Kan
A1 Emal Lesha
A1 Venkataramanan Soundararajan
A1 Feng Zhang
A1 George Church
YR 2016
UL http://biorxiv.org/content/early/2016/07/28/066597.abstract
AB Precise genetic modifications are essential for biomedical research and gene therapy. Yet, traditional homology-directed genome editing is limited by the requirements for DNA cleavage, donor DNA template and the endogenous DNA break-repair machinery. Here we present programmable cytidine deaminases that enable site-specific cytidine to thymidine (C-to-T) genomic edits without the need for DNA cleavage. Our targeted deaminases are efficient and specific in Escherichia coli, converting a genomic C-to-T with 13% efficiency and 95% accuracy. Edited cells do not harbor unintended genomic abnormalities. These novel enzymes also function in human cells, leading to a site-specific C-to-T transition in 2.5% of cells with reduced toxicity compared with zinc-finger nucleases. Targeted deaminases therefore represent a platform for safer and effective genome editing in prokaryotes and eukaryotes, especially in systems where DSBs are toxic, such as human stem cells and repetitive elements targeting.