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Quantum Dot-Based Molecular Beacons for Quantitative Detection of Nucleic Acids with CRISPR/Cas(N) Nucleases

ACS Nano. 2022-11; 
Christopher M Green, Joseph Spangler, Kimihiro Susumu, David A Stenger, Igor L Medintz, Sebastián A Díaz
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Catalog Peptides … The LwCas13a enzyme from Leptotrichia wadei was purchased from GenScript at a concentration of 37.6 μM in 50 mM Tris-HCl, 600 mM NaCl, 5% glycerol, and 2 mM DTT (pH 7.5) … Get A Quote

摘要

Strategies utilizing the CRISPR/Cas nucleases Cas13 and Cas12 have shown great promise in the development of highly sensitive and rapid diagnostic assays for the detection of pathogenic nucleic acids. The most common approaches utilizing fluorophore-quencher molecular beacons require strand amplification strategies or highly sensitive optical setups to overcome the limitations of the readout. Here, we demonstrate a flexible strategy for assembling highly luminescent and colorimetric quantum dot-nucleic acid hairpin (QD-HP) molecular beacons for use in CRISPR/Cas diagnostics. This strategy utilizes a chimeric peptide-peptide nucleic acid (peptide-PNA) to conjugate fluorescently labeled DNA or RNA hairpins to ZnS... More

关键词

CRISPR/Cas, Förster resonance energy transfer, Molecular beacons, Peptide nucleic acids, Quantum dots, Ratiometric