High-contrast imaging of cellular non-repetitive drug-resistant genes via in situ dead Cas12a-labeled PCR.
Chem Commun (Camb)
; 60(76): 10524-10527, 2024 Sep 20.
Article
em En
| MEDLINE
| ID: mdl-39229640
ABSTRACT
In situ imaging of genes of pathogenic bacteria can profile cellular heterogeneity, such as the emergence of drug resistance. Fluorescence in situ hybridization (FISH) serves as a classic approach to image mRNAs inside cells, but it remains challenging to elucidate genomic DNAs and relies on multiple fluorescently labeled probes. Herein, we present a dead Cas12a (dCas12a)-labeled polymerase chain reaction (CasPCR) assay for high-contrast imaging of cellular drug-resistant genes. We employed a syncretic dCas12a-green fluorescent protein (dCas12a-GFP) to tag the amplicons, thereby enabling high-contrast imaging and avoiding multiple fluorescently labeled probes. The CasPCR assay can quantify quinolone-resistant Salmonella enterica in mixed populations and identify them isolated from poultry farms.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
/
Salmonella enterica
Limite:
Animals
Idioma:
En
Revista:
Chem Commun (Camb)
/
Chem. commun. (Lond., 1996, Online)
/
Chemical communications (London. 1996. Online)
Assunto da revista:
QUIMICA
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
China
País de publicação:
Reino Unido