Chromatin accessibility is associated with CRISPR-Cas9 efficiency in the zebrafish (Danio rerio).

ABSTRACT: CRISPR-Cas9 technology is routinely applied for targeted mutagenesis in model organisms and cell lines. Recent studies indicate that the prokaryotic CRISPR-Cas9 system is affected by eukaryotic chromatin structures. Here, we show that the likelihood of successful mutagenesis correlates with transcript levels during early development in zebrafish (Danio rerio) embryos. In an experimental setting, we found that guide RNAs differ in their onset of mutagenesis activity in vivo. Furthermore, some guide RNAs with high in vitro activity possessed poor mutagenesis activity in vivo, suggesting the presence of factors that limit the mutagenesis in vivo. Using open access datasets generated from early developmental stages of the zebrafish, and guide RNAs selected from the CRISPRz database, we provide

SUBMITTER: Uusi-Makela MIE 

PROVIDER: S-EPMC5912780 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature