Programmable DNA Nanoswitch-Regulated Plasmonic CRISPR/Cas12a-Gold Nanostars Reporter Platform for Nucleic Acid and Non-Nucleic Acid Biomarker Analysis Assisted by a Spatial Confinement Effect

Nano Letters, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 15, 2025

CRISPR/Cas 12a system based nucleic acid and non-nucleic targets detection faces two challenges including (1) multiple crRNAs are needed for biomarkers (2) insufficient sensitivity resulted from photobleaching of fluorescent dyes the low kinetic cleavage rate a traditional single-strand (ssDNA) reporter. To address these limitations, we developed programmable DNA nanoswitch (NS)-regulated plasmonic CRISPR/Cas12a-gold nanostars (Au NSTs) reporter platform with assistance spatial confinement effect. Through simply programming target recognition sequence in NS, only one crRNA is required to detect both biomarkers. The limit decreased by ∼196-fold miRNA-375 122-fold prostate-specific antigen (PSA), respectively. Moreover, versatile evaluation PSA clinical urine samples can also be achieved, according which prostate cancer healthy groups well identified.

Language: Английский

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Phosphorothioate-Modified Hairpin G-Triplex Reporter-Assisted Split CRISPR/Cas12a-Powered Biosensor for “Turn-On” Fluorescent Detection of Nucleic Acid and Non-Nucleic Acid Targets

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 24, 2025

CRISPR/Cas12a-powered biosensors with guanine (G)-rich sequence reporters (e.g., G-quadruplex and G-triplex) are widely used in detection applications due to their simplicity sensitivity. However, when these employed for molecular complex samples, they may encounter difficulties such as high background signal susceptibility interference because of the "turn-off" output. Herein, we explore, first time, a set phosphorothioate (ps)-modified (G4) G-triplex (G3) sequences that can bind thioflavin T (ThT) an active split CRISPR/Cas12a system (SCas12a) generate "turn-on" fluorescent signal. To apply this new phenomenon, develop universal SCas12a-powered biosensor nucleic acid (miRNA-21) non-nucleic (kanamycin) targets by using ps-modified hairpin G3 reporter (SCas12a/psHG3). Target recognition activates SCas12a's trans-cleavage activity, leading cleavage at loop region psHG3 reporter. The released prelocked psG3 DNA binds ThT produce strong fluorescence Without preamplification, strategy detect miRNA-21 limit 100 fM. Moreover, SCas12a/psHG3 was further utilized detecting kanamycin incorporating its aptamers, enabling concentrations low pM. This work is system, showcasing improved performance wide range synthetic biology-based sensing technology.

Language: Английский

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CRISPR/Cas12a-Enabled Amplification-Free Colorimetric Visual Sensing Strategy for Point-of-Care Diagnostics of Biomarkers

Analytical Chemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 19, 2024

CRISPR/Cas12a-based biosensors have garnered significant attention in the field of point-of-care testing (POCT), yet majority CRISPR-based POCT methods employ fluorescent systems as report probes. Herein, we a new CRISPR/Cas12a-enabled multicolor visual biosensing strategy for rapid detection disease biomarkers. The proposed assay provided vivid color responses to enhance accuracy detection. In existence target,

Language: Английский

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CRISPR-Cas-based colorimetric strategies for nucleic acids detection

TrAC Trends in Analytical Chemistry, Journal Year: 2024, Volume and Issue: unknown, P. 118058 - 118058

Published: Nov. 1, 2024

Language: Английский

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