Analysing the outcome of CRISPR-aided genome editing in embryos: Screening, genotyping and quality control.
Methods
; 121-122: 68-76, 2017 05 15.
Article
em En
| MEDLINE
| ID: mdl-28363792
ABSTRACT
The application of CRISPR/Cas9 technology has revolutionised genetics by greatly enhancing the efficacy of genome editing in the early embryo. Furthermore, the system has enabled the generation of allele types previously incompatible with in vivo mutagenesis. Despite its versatility and ease of implementation, CRISPR/Cas9 editing outcome is unpredictable and can generate mosaic founders. Therefore, careful genotyping and characterisation of new mutants is proving essential. The literature presents a wide range of protocols for molecular characterisation, each representing different levels of investment. We present strategies and protocols for designing, producing and screening CRISPR/Cas9 edited founders and genotyping their offspring according to desired allele type (indel, point mutation and deletion).
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
RNA Guia de Cinetoplastídeos
/
Técnicas de Transferência de Genes
/
Endonucleases
/
Técnicas de Inativação de Genes
/
Sistemas CRISPR-Cas
/
Edição de Genes
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Limite:
Animals
Idioma:
En
Revista:
Methods
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Reino Unido