Programmable base editing in zebrafish using a modified CRISPR-Cas9 system

Publication date: Available online 2 August 2018Source: MethodsAuthor(s): Wei Qin, Xiaochan Lu, Shuo LinAbstractThe use of CRISPR/Cas9 to knockout genes in zebrafish has been well established. However, to better model many human diseases that are caused by point mutations, a robust methodology for generating desirable DNA base changes is still needed. Recently, Cas9-linked cytidine deaminases (base editors) evolved as a strategy to introduce single base mutations in model organisms. They have been used to convert cytidine to thymine at specific genomic loci. Here we describe a protocol for using the base editing system in zebrafish and its application to reproduce a single base mutation observed in human Ablepharon-Macrostomia Syndrome.
Source: Methods - Category: Molecular Biology Source Type: research