ZFIN ID: ZDB-PUB-141129-2
Generation of Zebrafish Models by CRISPR /Cas9 Genome Editing
Hruscha, A., Schmid, B.
Date: 2015
Source: Methods in molecular biology (Clifton, N.J.)   1254: 341-50 (Journal)
Registered Authors: Hruscha, Alexander, Schmid, Bettina
Keywords: none
MeSH Terms:
  • Animals
  • Caspase 9/genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics*
  • Genetic Engineering*
  • Genome
  • Molecular Biology/methods
  • Mutation
  • RNA Editing/genetics*
  • RNA, Guide/genetics
  • Zebrafish
PubMed: 25431076 Full text @ Meth. Mol. Biol.
The CRISPR /Cas system identified in archaea has been adopted and optimized for genome editing purposes in zebrafish. In vitro transcribed guide RNA and Cas9 mRNA are microinjected into fertilized zebrafish embryos to edit the zebrafish genome. Here, we describe how to design a gRNA , a fast method for in vitro transcription of gRNA from oligonucleotides , microinjection into fertilized zebrafish embryos, and a PCR -based restriction fragment length assay to identify mutations at the gRNA target site.