ZFIN ID: ZDB-PUB-181227-6
Fluorescently Labeled TracrRNA Improves Work Flow and Facilitates Successful Genome Editing in Zebrafish
Hamimi, M., Khabooshan, M., Castillo, H.A., Kaslin, J.
Date: 2018
Source: Zebrafish   16(1): 135-137 (Other)
Registered Authors: Kaslin, Jan
Keywords: CRISPR, knockin, knockout, loss-of-function, mutant, teleost
MeSH Terms:
  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Gene Editing/instrumentation
  • Gene Editing/methods*
  • Genome*
  • Heterocyclic Compounds, 4 or More Rings/chemistry*
  • Zebrafish/genetics*
PubMed: 30585775 Full text @ Zebrafish
Gene editing using clustered regularly interspaced short palindromic repeats (CRISPR) is widely used throughout the zebrafish community for the generation of knockouts and knockins. One of the bottlenecks that exists during the process is the laborious screening of injected embryos for F0 founder fish or CRISPants, weeks after the injection date. In this study we show that the use of fluorescently tagged tracrRNA and sorting for fluorescent embryos as early as the 512-cell stage using stereomicroscope significantly improve yield of fish with successfully CRISPR/Cas9-edited genomes. This is a cost-effective strategy that significantly improves workflow and efficacy in genome editing in particular for less experienced researchers.