Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair

• DOI: 10.7554/elife.33761
• Publication Date: 2018-05-29T12:01:43Z
• Authors: Natasa Savic; Femke CAS Ringnalda; Helen Lindsay; Christian Berk; Katja Bargsten; Yizhou Li; Dario Neri; Mark D Robinson; Constance Ciaudo; Jonathan Hall; Martin Jinek; Gerald Schwank
• Keywords: Nuclease; Homology directed repair; Coding strand

The CRISPR-Cas9 targeted nuclease technology allows the insertion of genetic modifications with single base-pair precision. preference mammalian cells to repair Cas9-induced DNA double-strand breaks via error-prone end-joining pathways rather than homology-directed mechanisms, however, leads relatively low rates precise editing from donor DNA. Here we show that spatial and temporal co-localization template Cas9 covalent linkage increases correction up 24-fold, demonstrate effect is mainly caused by an increase concentration in nucleus. Enhanced were observed multiple cell types on different genomic loci, suggesting covalently linking complex provides advantages for clinical applications where high-fidelity desired.