PUBLICATION

An Accessible Protocol for the Generation of CRISPR-Cas9 Knockouts Using INDELs in Zebrafish

Authors
Moravec, C.E., Pelegri, F.J.
ID
ZDB-PUB-190210-11
Date
2019
Source
Methods in molecular biology (Clifton, N.J.)   1920: 377-392 (Chapter)
Registered Authors
Pelegri, Francisco
Keywords
CRISPR-Cas9, Genome editing, INDELS, Targeted mutations, Zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • CRISPR-Cas Systems*
  • Computational Biology/methods
  • Female
  • Gene Editing*
  • Gene Knockout Techniques*
  • Gene Targeting*/methods
  • INDEL Mutation*
  • Male
  • RNA, Guide/chemical synthesis
  • RNA, Guide/genetics
  • Zebrafish/genetics*
PubMed
30737704 Full text @ Meth. Mol. Biol.
Abstract
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
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes