Tracking of Indels by DEcomposition is a Simple and Effective Method to Assess Efficiency of Guide RNAs in Zebrafish.
Zebrafish
; 14(6): 586-588, 2017 12.
Article
em En
| MEDLINE
| ID: mdl-28767326
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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Peixe-Zebra
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Técnicas Genéticas
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RNA Guia de Cinetoplastídeos
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Mutação INDEL
Tipo de estudo:
Evaluation_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article