RT Journal Article
SR Electronic
T1 Enabling large-scale genome editing by reducing DNA nicking
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 574020
DO 10.1101/574020
A1 Cory J. Smith
A1 Oscar Castanon
A1 Khaled Said
A1 Verena Volf
A1 Parastoo Khoshakhlagh
A1 Amanda Hornick
A1 Raphael Ferreira
A1 Chun-Ting Wu
A1 Marc Güell
A1 Shilpa Garg
A1 Hannu Myllykallio
A1 George M. Church
YR 2019
UL http://biorxiv.org/content/early/2019/03/15/574020.abstract
AB To extend the frontier of genome editing and enable the radical redesign of mammalian genomes, we developed a set of dead-Cas9 base editor (dBEs) variants that allow editing at tens of thousands of loci per cell by overcoming the cell death associated with DNA double-strand breaks (DSBs) and single-strand breaks (SSBs). We used a set of gRNAs targeting repetitive elements – ranging in target copy number from about 31 to 124,000 per cell. dBEs enabled survival after large-scale base editing, allowing targeted mutations at up to ~13,200 and ~2610 loci in 293T and human induced pluripotent stem cells (hiPSCs), respectively, three orders of magnitude greater than previously recorded. These dBEs can overcome current on-target mutation and toxicity barriers that prevent cell survival after large-scale genome engineering.One Sentence Summary Base editing with reduced DNA nicking allows for the simultaneous editing of >10,000 loci in human cells.