An efficient system for homology-dependent targeted gene integration in medaka (Oryzias latipes).

ABSTRACT:

Background

The CRISPR/Cas system is a powerful genome editing tool that enables targeted genome modifications in various organisms. In medaka (Oryzias latipes), targeted mutagenesis with small insertions and deletions using this system have become a robust technique and are now widely used. However, to date there have been only a small number of reports on targeted gene integration using this system. We thus sought in the present study to identify factors that enhance the efficiency of targeted gene integration events in medaka.

Results

We show that longer homology arms (ca. 500 bp) and linearization of circular donor plasmids by cleavage with bait sequences enhances the efficiency of targeted integration of plasmids in embryos. A new bait sequence, BaitD, which we de

SUBMITTER: Murakami Y 

PROVIDER: S-EPMC5500998 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature