PUBLICATION
Generation of Targeted Genomic Deletions Through CRISPR/Cas System in Zebrafish
- Authors
- Xiao, A., Zhang, B.
- ID
- ZDB-PUB-160729-25
- Date
- 2016
- Source
- Methods in molecular biology (Clifton, N.J.) 1451: 65-79 (Chapter)
- Registered Authors
- Xiao, An, Zhang, Bo
- Keywords
- CRISPR/Cas, Chromosomal deletion, Gene disruption, Gene targeting, Genome manipulation, TALEN, Zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems/genetics*
- Chromosome Deletion
- Clustered Regularly Interspaced Short Palindromic Repeats/genetics
- Genetic Engineering
- Genome/genetics*
- INDEL Mutation/genetics
- RNA, Guide, Kinetoplastida/genetics
- Sequence Deletion/genetics*
- Zebrafish/genetics
- PubMed
- 27464801 Full text @ Meth. Mol. Biol.
Citation
Xiao, A., Zhang, B. (2016) Generation of Targeted Genomic Deletions Through CRISPR/Cas System in Zebrafish. Methods in molecular biology (Clifton, N.J.). 1451:65-79.
Abstract
Using TALEN or CRISPR/Cas system to induce small indels into coding sequences has been implicated in broad applications for reverse genetic studies of many organisms including zebrafish. However, complete deletion of a large gene or noncoding gene(s) or removing a large genomic fragment spanning several genes or other chromosomal elements is preferred in various cases, as well as inducing chromosomal inversions. Here, we describe the detailed protocols for the generation of chromosomal deletion mutations mediated by Cas9 and a pair of gRNAs and the evaluation for the efficiencies in F0 founder fish and of germline transmission.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping