PUBLICATION
Detection of Multiple Genome Modifications Induced by the CRISPR/Cas9 System
- Authors
- Ota, S., Kawahara, A.
- ID
- ZDB-PUB-160729-26
- Date
- 2016
- Source
- Methods in molecular biology (Clifton, N.J.) 1451: 53-63 (Chapter)
- Registered Authors
- Kawahara, Atsuo, Ota, Satoshi
- Keywords
- CRISPR/Cas9, Genome modifications, HMA, Multi-locus HMA, Zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems/genetics*
- Clustered Regularly Interspaced Short Palindromic Repeats/genetics
- Genetic Engineering
- Genome/genetics*
- Mutation
- RNA, Guide, Kinetoplastida/genetics
- Zebrafish/genetics
- PubMed
- 27464800 Full text @ Meth. Mol. Biol.
Citation
Ota, S., Kawahara, A. (2016) Detection of Multiple Genome Modifications Induced by the CRISPR/Cas9 System. Methods in molecular biology (Clifton, N.J.). 1451:53-63.
Abstract
The recent remarkable innovation of an RNA-guided nuclease system, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, enables us the modification of specific genomic loci in various model animals including zebrafish. With this system, multiple guide RNAs simultaneously injected with the Cas9 nuclease into zebrafish embryos cause multiple genome modifications at different genomic loci with high efficiency; therefore, a simple method to detect individual mutations at distinct loci is desired. In this chapter, we describe a procedure for inducing multiple CRISPR/Cas9-mediated genome modifications in zebrafish and a convenient method to detect CRISPR/Cas9-induced insertion and/or deletion (indel) mutations using a heteroduplex mobility assay (HMA).
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping