ZFIN ID: ZDB-PUB-170628-4
RICE CRISPR: Rapidly Increased Cut Ends by an Exonuclease Cas9 Fusion in Zebrafish
Clements, T.P., Tandon, B., Lintel, H.A., McCarty, J.H., Wagner, D.S.
Date: 2017
Source: Genesis (New York, N.Y. : 2000) 55(8): (Journal)
Registered Authors: Wagner, Daniel
Keywords: Danio rerio, S. pyogenes Cas9, Targeted Mutagenesis
MeSH Terms:
  • Animals
  • Bacterial Proteins/genetics*
  • Bacterial Proteins/metabolism
  • CRISPR-Cas Systems*
  • Endonucleases/genetics*
  • Endonucleases/metabolism
  • Gene Deletion
  • Gene Targeting/methods*
  • Gene Targeting/standards
  • Loss of Function Mutation
  • Zebrafish/genetics*
PubMed: 28653435 Full text @ Genesis
ABSTRACT
Application of CRISPR-Cas9 technology in diverse organisms has resulted in an explosion of genome modification efforts. To expand the toolbox of applications, we have created an E. coli Exonuclease I (sbcB) - Cas9 fusion that has altered enzymatic activity in zebrafish embryos. This Cas9 variant has increased mutation efficiency and favors longer deletions relative to wild type Cas9. We anticipate that this variant will allow for more efficient screening for F0 phenotypes and mutation of a larger spectrum of genomic targets including deletion of regulatory regions and creating loss of function mutations in transcription units with poor sequence conservation such as lncRNAs where larger deletions may be required for loss of function. This article is protected by copyright. All rights reserved.
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