Exploring the Effect of Steric Hindrance on Trans-cleavage Activity of CRISPR-cas12a for Ultrasensitive SERS Detection of P53 DNA.
Anal Chem
; 96(26): 10654-10661, 2024 07 02.
Article
em En
| MEDLINE
| ID: mdl-38875020
ABSTRACT
The trans-cleavage properties of Cas12a make it important for gene editing and disease diagnosis. In this work, the effect of spatial site resistance on the trans-cleavage activity of Cas12a was studied. First, we have explored the cutting effect of Cas12a when different-sized nanoparticles are linked with various spacings of DNA strands using the fluorescence method. The minimum spacing with different-sized nanoparticles that cas12a can cut was determined. We found that when the size of the nanoparticles increases, the minimum spacing that cas12a can cut gradually increases. Subsequently, we verified the conclusion using the surface-enhanced Raman scattering (SERS) method, and at the same time, we designed a SERS biosensor that can achieve ultrasensitive detection of P53 DNA with a linear range of 1 fM-10 nM and a limit of detection of 0.40 fM. Our work develops a deep study of the trans-cleavage activity of Cas12a and gives a guide for DNA design in cas12a-related studies, which can be applied in biomedical analysis and other fields.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Análise Espectral Raman
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DNA
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Proteína Supressora de Tumor p53
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Sistemas CRISPR-Cas
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article