ZFIN ID: ZDB-PUB-170803-3
Tracking of Indels by DEcomposition is a Simple and Effective Method to Assess Efficiency of Guide RNAs in Zebrafish
Etard, C., Joshi, S., Stegmaier, J., Mikut, R., Strähle, U.
Date: 2017
Source: Zebrafish   14(6): 586-588 (Journal)
Registered Authors: Etard, Christelle, Joshi, Swarnima, Mikut, Ralf, Strähle, Uwe
Keywords: TIDE, efficiencies, guide-RNAs, zebrafish
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems
  • Gene Editing
  • Genetic Techniques/veterinary*
  • INDEL Mutation*
  • Polymerase Chain Reaction/methods
  • RNA, Guide/genetics*
  • Sequence Analysis, RNA/methods
  • Zebrafish/genetics*
PubMed: 28767326 Full text @ Zebrafish
ABSTRACT
A bottleneck in CRISPR/Cas9 genome editing is variable efficiencies of in silico-designed gRNAs. We evaluated the sensitivity of the TIDE method (Tracking of Indels by DEcomposition) introduced by Brinkman et al. in 2014 for assessing the cutting efficiencies of gRNAs in zebrafish. We show that this simple method, which involves bulk polymerase chain reaction amplification and Sanger sequencing, is highly effective in tracking well-performing gRNAs in pools of genomic DNA derived from injected embryos. The method is equally effective for tracing INDELs in heterozygotes.
ADDITIONAL INFORMATION No data available