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Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
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Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
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Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
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Journal Article
Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System
2016
Overview
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.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Amyotrophic lateral sclerosis
/ Amyotrophic Lateral Sclerosis - genetics
/ Analysis
/ Animals
/ CRISPR
/ CRISPR-Cas Systems - genetics
/ DNA-Binding Proteins - genetics
/ Editing
/ Gene Knock-In Techniques - methods
/ Genes
/ Genomes
/ Homology
/ Humans
/ Mutation
/ Oligodeoxyribonucleotides - genetics
/ Polymorphism, Single Nucleotide
/ Research and Analysis Methods
/ RNA-Binding Protein FUS - genetics
/ Rodents
/ Sequence Homology, Amino Acid
/ Sequence Homology, Nucleic Acid
