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 DOI
Kai Shi,

Wenjie Luo,

Ying Cheng

et al.

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

A Critical Review of Sensors for Detecting Per- and Polyfluoroalkyl Substances: Focusing on Diverse Molecular Probes DOI

Jiancheng Zha,

Muyuan Ma,

Yue Shen

et al.

Environmental Research, Journal Year: 2025, Volume and Issue: unknown, P. 121669 - 121669

Published: April 1, 2025

Language: Английский

Citations

0
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 DOI
Kai Shi,

Wenjie Luo,

Ying Cheng

et al.

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

Citations

0