Hydrazone chemistry-mediated CRISPR/Cas12a system for bacterial analysis.
Nucleic Acids Res
; 50(18): 10562-10570, 2022 10 14.
Article
em En
| MEDLINE
| ID: mdl-36156138
ABSTRACT
In this study, a hydrazone chemistry-mediated clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 12a (Cas12a) system has been proposed for the fist time and constructed. In our system, hydrazone chemistry is designed and employed to accelerate the formation of a whole activation strand by taking advantage of the proximity effect induced by complementary base pairing, thus activating the CRISPR/Cas12a system quickly and efficiently. Moreover, the introduction of hydrazone chemistry can improve the specificity of the CRISPR/Cas12a system, allowing it to effectively distinguish single-base mismatches. The established system has been further applied to analyze Pseudomonas aeruginosa by specific recognition of the probe strand with a characteristic fragment in 16S rDNA to release the hydrazine group-modified activation strand. The method shows a wide linear range from 3.8 × 102 colony-forming units (CFU)/ml to 3.8 × 106 CFU/ml, with the lowest detection limit of 24 CFU/ml. Therefore, the introduction of hydrazone chemistry may also broaden the application of the CRISPR/Cas12a system.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Associadas a CRISPR
/
Sistemas CRISPR-Cas
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article