PUBLICATION
Generation of Zebrafish Models by CRISPR /Cas9 Genome Editing
- Authors
- Hruscha, A., Schmid, B.
- ID
- ZDB-PUB-141129-2
- Date
- 2015
- Source
- Methods in molecular biology (Clifton, N.J.) 1254: 341-50 (Journal)
- Registered Authors
- Hruscha, Alexander, Schmid, Bettina
- Keywords
- none
- MeSH Terms
-
- RNA Editing/genetics*
- Genetic Engineering*
- Zebrafish
- Genome
- RNA, Guide, Kinetoplastida/genetics
- Mutation
- Caspase 9/genetics*
- Molecular Biology/methods
- Animals
- Clustered Regularly Interspaced Short Palindromic Repeats/genetics*
- PubMed
- 25431076 Full text @ Meth. Mol. Biol.
Citation
Hruscha, A., Schmid, B. (2015) Generation of Zebrafish Models by CRISPR /Cas9 Genome Editing. Methods in molecular biology (Clifton, N.J.). 1254:341-50.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping