PUBLICATION
Multiplex conditional mutagenesis in zebrafish using the CRISPR/Cas system
- Authors
- Yin, L., Maddison, L.A., Chen, W.
- ID
- ZDB-PUB-160725-35
- Date
- 2016
- Source
- Methods in cell biology 135: 3-17 (Chapter)
- Registered Authors
- Chen, Wenbiao
- Keywords
- CRISPR, Cas9, Conditional mutagenesis, Mutagenesis, Transgenesis, sgRNA
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- CRISPR-Cas Systems/genetics*
- Gene Editing/methods*
- Gene Transfer Techniques
- Genetic Engineering/methods*
- Mutagenesis/genetics*
- Mutation/genetics
- Zebrafish/genetics
- Zebrafish/growth & development
- PubMed
- 27443918 Full text @ Meth. Cell. Biol.
Citation
Yin, L., Maddison, L.A., Chen, W. (2016) Multiplex conditional mutagenesis in zebrafish using the CRISPR/Cas system. Methods in cell biology. 135:3-17.
Abstract
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system is a powerful tool for genome editing in numerous organisms. However, the system is typically used for gene editing throughout the entire organism. Tissue and temporal specific mutagenesis is often desirable to determine gene function in a specific stage or tissue and to bypass undesired consequences of global mutations. We have developed the CRISPR/Cas system for conditional mutagenesis in transgenic zebrafish using tissue-specific and/or inducible expression of Cas9 and U6-driven expression of sgRNA. To allow mutagenesis of multiple targets, we have isolated four distinct U6 promoters and designed Golden Gate vectors to easily assemble transgenes with multiple sgRNAs. We provide experimental details on the reagents and applications for multiplex conditional mutagenesis in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping