An Accessible Protocol for the Generation of CRISPR-Cas9 Knockouts Using INDELs in Zebrafish
- Moravec, C.E., Pelegri, F.J.
- Methods in molecular biology (Clifton, N.J.) 1920: 377-392 (Chapter)
- Registered Authors
- Pelegri, Francisco
- CRISPR-Cas9, Genome editing, INDELS, Targeted mutations, Zebrafish
- MeSH Terms
- Animals, Genetically Modified
- CRISPR-Cas Systems*
- Computational Biology/methods
- Gene Editing*
- Gene Knockout Techniques*
- Gene Targeting*/methods
- INDEL Mutation*
- RNA, Guide/chemical synthesis
- RNA, Guide/genetics
- 30737704 Full text @ Meth. Mol. Biol.
Moravec, C.E., Pelegri, F.J. (2019) An Accessible Protocol for the Generation of CRISPR-Cas9 Knockouts Using INDELs in Zebrafish. Methods in molecular biology (Clifton, N.J.). 1920:377-392.
The ability to create targeted mutations in specific genes, and therefore a loss-of-function condition, provides essential information about their endogenous functions during development and homeostasis. The discovery that CRISPR-Cas9 can target specific sequences according to base-pair complementarity and readily create knockouts in a desired gene has elevated the implementation of genetic analysis in numerous organisms. As CRISPR-Cas9 has become a powerful tool in a number of species, multiple methods for designing, creating, and screening editing efficiencies have been published, each of which has unique benefits. This chapter presents a cost-efficient, accessible protocol for creating knockout mutants in zebrafish using insertions/deletions (INDELS), from target site selection to mutant propagation, using basic laboratory supplies. The presented approach can be adapted to other systems, including any vertebrate species.
Genes / Markers
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes