|ZFIN ID: ZDB-PUB-160302-4|
Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
Armstrong, G.A., Liao, M., You, Z., Lissouba, A., Chen, B.E., Drapeau, P.
|Source:||PLoS One 11: e0150188 (Journal)|
|Registered Authors:||Drapeau, Pierre|
|Keywords:||Point mutation, Zebrafish, Sequence motif analysis, Non-homologous end joining, Polymerase chain reaction, Embryos, Missense mutation, Restriction fragment length polymorphism analysis|
|PubMed:||26930076 Full text @ PLoS One|
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.
ABSTRACTThe 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.