PUBLICATION

CRISPR/Cas9-Mediated Targeted Knockin of Exogenous Reporter Genes in Zebrafish

Authors
Kawahara, A.
ID
ZDB-PUB-170624-4
Date
2017
Source
Methods in molecular biology (Clifton, N.J.)   1630: 165-173 (Chapter)
Registered Authors
Kawahara, Atsuo
Keywords
CRISPR/Cas9, HR, Knockin, Knockout, MMEJ, NHEJ, Zebrafish
MeSH Terms
  • Animals
  • CRISPR-Cas Systems
  • Frameshift Mutation
  • Gene Editing/methods*
  • Gene Knock-In Techniques
  • Genes, Reporter*
  • Recombination, Genetic
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed
28643258 Full text @ Meth. Mol. Biol.
Abstract
Genome editing technologies such as ZFN, TALEN, and CRISPR/Cas9 efficiently induce DNA double-stranded breaks (DSBs) at a targeted genomic locus, often resulting in a frameshift-mediated target gene disruption. It remains difficult to perform targeted integration of exogenous genes by genome editing technologies. DSBs can be restored through DNA repair mechanisms, such as non-homologous end joining (NHEJ), microhomology-mediated end joining (MMEJ), and homologous recombination (HR). It is well known that HR facilitates homology-dependent integration of donor DNA template into a targeted locus. Recently, both NHEJ-mediated and MMEJ-mediated targeted integrations of exogenous genes have been developed in zebrafish. This chapter summarizes the application of CRISPR/Cas9-mediated knock-in technology in zebrafish.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping