CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity.
Science
; 360(6387): 436-439, 2018 04 27.
Article
em En
| MEDLINE
| ID: mdl-29449511
ABSTRACT
CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems. Like CRISPR-Cas9, Cas12a has been harnessed for genome editing on the basis of its ability to generate targeted, double-stranded DNA breaks. Here we show that RNA-guided DNA binding unleashes indiscriminate single-stranded DNA (ssDNA) cleavage activity by Cas12a that completely degrades ssDNA molecules. We find that target-activated, nonspecific single-stranded deoxyribonuclease (ssDNase) cleavage is also a property of other type V CRISPR-Cas12 enzymes. By combining Cas12a ssDNase activation with isothermal amplification, we create a method termed DNA endonuclease-targeted CRISPR trans reporter (DETECTR), which achieves attomolar sensitivity for DNA detection. DETECTR enables rapid and specific detection of human papillomavirus in patient samples, thereby providing a simple platform for molecular diagnostics.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
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DNA de Cadeia Simples
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Endonucleases
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Clivagem do DNA
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Proteínas Associadas a CRISPR
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Sistemas CRISPR-Cas
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Clostridiales
Idioma:
En
Revista:
Science
Ano de publicação:
2018
Tipo de documento:
Article