Journal
ZEBRAFISH
Volume 14, Issue 6, Pages 586-588Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/zeb.2017.1454
Keywords
guide-RNAs; efficiencies; zebrafish; TIDE
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Funding
- HGF
- DFG [STR 39/8-1]
- BMBF-MIE Molecular Interaction Engineering'' [031A095 C]
- EU Horizon 2020, Marie Sklodowska-Curie program [643062]
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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.
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