Efficient single-copy HDR by 5' modified long dsDNA donors

Jose Arturo Gutierrez-Triana, Tinatini Tavhelidse, Thomas Thumberger, Isabelle Thomas, Beate Wittbrodt, Tanja Kellner, Kerim Anlas, Erika Tsingos, Joachim Wittbrodt
2018 eLife  
CRISPR/Cas9 efficiently induces targeted mutations via non-homologous-end-joining but for genome editing, precise, homology-directed repair (HDR) of endogenous DNA stretches is a prerequisite. To favor HDR, many approaches interfere with the repair machinery or manipulate Cas9 itself. Using Medaka we show that the modification of 5' ends of long dsDNA donors strongly enhances HDR, favors efficient single-copy integration by retaining a monomeric donor conformation thus facilitating successful
more » ... tating successful gene replacement or tagging.
doi:10.7554/elife.39468 pmid:30156184 fatcat:5a7mya5a7fftfja5ibi7p6kg6q