ZFIN ID: ZDB-PUB-130805-29
Heritable and Precise Zebrafish Genome Editing Using a CRISPR-Cas System
Hwang, W.Y., Fu, Y., Reyon, D., Maeder, M.L., Kaini, P., Sander, J.D., Joung, J.K., Peterson, R.T., and Yeh, J.R.
Date: 2013
Source: PLoS One   8(7): e68708 (Journal)
Registered Authors: Peterson, Randall, Yeh, Jing-Ruey (Joanna)
Keywords: none
MeSH Terms:
  • Animals
  • Base Sequence
  • CRISPR-Associated Proteins/chemistry
  • CRISPR-Associated Proteins/genetics
  • CRISPR-Cas Systems*
  • Genetic Engineering
  • Genome*
  • Germ-Line Mutation
  • INDEL Mutation
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Mutation Rate
  • Oligonucleotides/chemistry
  • Oligonucleotides/genetics
  • Sequence Alignment
  • Zebrafish/genetics*
PubMed: 23874735 Full text @ PLoS One

We have previously reported a simple and customizable CRISPR (clustered regularly interspaced short palindromic repeats) RNA-guided Cas9 nuclease (RGN) system that can be used to efficiently and robustly introduce somatic indel mutations in endogenous zebrafish genes. Here we demonstrate that RGN-induced mutations are heritable, with efficiencies of germline transmission reaching as high as 100%. In addition, we extend the power of the RGN system by showing that these nucleases can be used with single-stranded oligodeoxynucleotides (ssODNs) to create precise intended sequence modifications, including single nucleotide substitutions. Finally, we describe and validate simple strategies that improve the targeting range of RGNs from 1 in every 128 basepairs (bps) of random DNA sequence to 1 in every 8 bps. Together, these advances expand the utility of the CRISPR-Cas system in the zebrafish beyond somatic indel formation to heritable and precise genome modifications.