ZFIN ID: ZDB-PUB-181028-2
Zebrafish knock-ins swim into the mainstream
Prykhozhij, S.V., Berman, J.N.
Date: 2018
Source: Disease models & mechanisms   11(10): (Review)
Registered Authors: Berman, Jason, Prykhozhij, Sergey
Keywords: CRISPR/Cas9, Genome editing, Point mutations, Zebrafish
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems/genetics
  • Disease Models, Animal
  • Gene Knock-In Techniques*
  • Point Mutation/genetics
  • Zebrafish/genetics*
PubMed: 30366936 Full text @ Dis. Model. Mech.
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ABSTRACT
The zebrafish is an increasingly popular model organism for human genetic disease research. CRISPR/Cas9-based approaches are currently used for multiple gene-editing purposes in zebrafish, but few studies have developed reliable ways to introduce precise mutations. Point mutation knock-in using CRISPR/Cas9 and single-stranded oligodeoxynucleotides (ssODNs) is currently the most promising technology for this purpose. Despite some progress in applying this technique to zebrafish, there is still a great need for improvements in terms of its efficiency, optimal design of sgRNA and ssODNs and broader applicability. The papers discussed in this Editorial provide excellent case studies on identifying problems inherent in the mutation knock-in technique, quantifying these issues and proposing strategies to overcome them. These reports also illustrate how the procedures for introducing specific mutations can be straightforward, such that ssODNs with only the target mutation are sufficient for generating the intended knock-in animals. Two of the studies also develop interesting point mutant knock-in models for cardiac diseases, validating the translational relevance of generating knock-in mutations and opening the door to many possibilities for their further study.
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