A single blastocyst assay optimized for detecting CRISPR/Cas9 system-induced indel mutations in mice.

ABSTRACT:

Background

Microinjection of clustered regulatory interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-related RNA and DNA into fertilized eggs is a novel approach for creating gene-modified mice. Blastocysts obtained just before implantation may be appropriate for testing the fidelity of CRIPSR/Cas9-mediated genome editing because they can be individually handled in vitro and obtained 3days after microinjection, thus allowing researchers to check mutations rapidly. However, it is not known whether indel mutations caused by the CRISPR/Cas9 system can be reproducibly detected in embryos. In this study, we assessed the detection of CRISPR/Cas9-induced mutations in embryos.

Results

T7 endonuclease I was more effective than Surveyor nuclease for detec

SUBMITTER: Sakurai T 

PROVIDER: S-EPMC4118159 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature