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PLoS One. 2016 Mar 1;11(3):e0150188. doi: 10.1371/journal.pone.0150188. eCollection 2016.

Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System.

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Department of Neurosciences, Research Centre of the University of Montréal Hospital Centre, Montréal, Québec, Canada.
Department of Neurology and Neurosurgery, Research Institute of the McGill University Health Centre and Centre for Research in Neuroscience, Montréal, Québec, Canada.


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.

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