nl0c04303_si_001.pdf (996.06 kB)
CRISPR-Cas12a-Based Nucleic Acid Amplification-Free DNA Biosensor via Au Nanoparticle-Assisted Metal-Enhanced Fluorescence and Colorimetric Analysis
journal contribution
posted on 2020-12-21, 14:38 authored by Jin-Ha Choi, Joungpyo Lim, Minkyu Shin, Se-Hwan Paek, Jeong-Woo ChoiCell-free
DNA (cfDNA) has attracted significant attention due to
its high potential to diagnose diseases, such as cancer. Still, its
detection by amplification method has limitations because of false-positive
signals and difficulty in designing target-specific primers. CRISPR-Cas-based
fluorescent biosensors have been developed but also need the amplification
step for the detection. In this study, for the first time CRISPR-Cas12a
based nucleic acid amplification-free fluorescent biosensor was developed
to detect cfDNA by a metal-enhanced fluorescence (MEF) using DNA-functionalized
Au nanoparticle (AuNP). Upon activating the CRISPR-Cas12a complex
by the target cfDNA and subsequent single-strand DNA (ssDNA) degradation
between AuNP and fluorophore, MEF occurred with color changes from
purple to red-purple. Using this system, breast cancer gene-1 (BRCA-1)
can be detected with very high sensitivity in 30 min. This rapid and
highly selective sensor can be applied to measure other nucleic acid
biomarkers such as viral DNA in field-deployable and point-of-care
testing (POCT) platform.
History
Usage metrics
Categories
Keywords
point-of-care testingmetal-enhanced fluorescencefalse-positive signalsdetectionsingle-strand DNAtarget-specific primersacid biomarkersbreast cancer geneAmplification-Free DNA BiosensorPOCTcolor changesCRISPR-Cas 12a Nucleicamplification stepCRISPR-Cas 12atime CRISPR-Cas 12aacid amplification-freeAuNPamplification methodbiosensorMEFNanoparticle-Assisted Metal-Enhance...Colorimetric Analysistarget cfDNABRCA30 min