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.
Citation
Kawahara, A. (2017) CRISPR/Cas9-Mediated Targeted Knockin of Exogenous Reporter Genes in Zebrafish. Methods in molecular biology (Clifton, N.J.). 1630:165-173.
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
Probes
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping