@article{54680e84b3fb451dbba882e898b22ae9,
title = "Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair",
abstract = "The CRISPR-Cas9 targeted nuclease technology allows the insertion of genetic modifications with single base-pair precision. The preference of mammalian cells to repair Cas9-induced DNA double-strand breaks via error-prone end-joining pathways rather than via homology-directed repair mechanisms, however, leads to relatively low rates of precise editing from donor DNA. Here we show that spatial and temporal co-localization of the donor template and Cas9 via covalent linkage increases the correction rates up to 24-fold, and demonstrate that the effect is mainly caused by an increase of donor template concentration in the nucleus. Enhanced correction rates were observed in multiple cell types and on different genomic loci, suggesting that covalently linking the donor template to the Cas9 complex provides advantages for clinical applications where high-fidelity repair is desired.",
keywords = "CRISPR-Associated Protein 9/genetics, CRISPR-Cas Systems, Cell Nucleus/genetics, Clustered Regularly Interspaced Short Palindromic Repeats, DNA/chemistry, DNA Breaks, Double-Stranded, DNA End-Joining Repair, DNA Replication, Gene Editing/methods, Genetic Loci, Guanidines/chemistry, HEK293 Cells, Humans, Oligodeoxyribonucleotides/chemistry, RNA, Guide, CRISPR-Cas Systems/genetics, Recombinant Fusion Proteins/genetics, Recombinational DNA Repair, Staining and Labeling/methods",
author = "Natasa Savic and Ringnalda, \{Femke Cas\} and Helen Lindsay and Christian Berk and Katja Bargsten and Yizhou Li and Dario Neri and Robinson, \{Mark D\} and Constance Ciaudo and Jonathan Hall and Martin Jinek and Gerald Schwank",
note = "{\textcopyright} 2018, Savic et al.",
year = "2018",
month = may,
day = "29",
doi = "10.7554/eLife.33761",
language = "English",
volume = "7",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications Ltd",
}