PUBLICATION
Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
- Authors
- Armstrong, G.A., Liao, M., You, Z., Lissouba, A., Chen, B.E., Drapeau, P.
- ID
- ZDB-PUB-160302-4
- Date
- 2016
- Source
- PLoS One 11: e0150188 (Journal)
- Registered Authors
- Armstrong, Gary A.B., Drapeau, Pierre
- Keywords
- Point mutation, Zebrafish, Sequence motif analysis, Non-homologous end joining, Polymerase chain reaction, Embryos, Missense mutation, Restriction fragment length polymorphism analysis
- MeSH Terms
-
- Amino Acid Sequence
- Amyotrophic Lateral Sclerosis/genetics*
- Animals
- Base Sequence
- CRISPR-Cas Systems/genetics*
- DNA Repair
- DNA, Single-Stranded
- DNA-Binding Proteins/genetics*
- Disease Models, Animal
- Gene Knock-In Techniques/methods
- Humans
- Oligodeoxyribonucleotides/genetics
- Point Mutation*
- Polymorphism, Single Nucleotide
- RNA-Binding Protein FUS/genetics*
- Reproducibility of Results
- Sequence Homology, Amino Acid
- Sequence Homology, Nucleic Acid
- Zebrafish/genetics
- Zebrafish Proteins/genetics*
- PubMed
- 26930076 Full text @ PLoS One
Citation
Armstrong, G.A., Liao, M., You, Z., Lissouba, A., Chen, B.E., Drapeau, P. (2016) Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System. PLoS One. 11:e0150188.
Abstract
The methodology for site-directed editing of single nucleotides in the vertebrate genome is of considerable interest for research in biology and medicine. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 type II (Cas9) system has emerged as a simple and inexpensive tool for editing genomic loci of interest in a variety of animal models. In zebrafish, error-prone non-homologous end joining (NHEJ) has been used as a simple method to disrupt gene function. We sought to develop a method to easily create site-specific SNPs in the zebrafish genome. Here, we report simple methodologies for using CRISPR/Cas9-mediated homology directed repair using single-stranded oligodeoxynucleotide donor templates (ssODN) for site-directed single nucleotide editing, for the first time in two disease-related genes, tardbp and fus.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping