CRISPR/Cas12a-Powered Electrochemical Platform for Dual-miRNA Detection via an AND Logic Circuit DOI

Jiangbo Dong,

Changjun Hou, Liyuan Deng

et al.

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

Published: Dec. 24, 2024

The CRISPR/Cas technology shows great potential in molecular detection and diagnostics. However, it is still challenging to detect multiple targets simultaneously using the CRISPR-Cas system. Herein, we ingeniously leverage synergistic effect of two short single-stranded DNA activators construct a CRISPR/Cas12a-driven electrochemical sensing platform based on an AND logic circuit ("AND" LC-CRISPR) for simultaneous dual miRNAs. Specifically, exponential amplification reaction products triggered by dual-specific miRNAs are designed as binary inputs bind with Cas12a/crRNA, forming activating trans-cleavage ability CRISPR-Cas12a Subsequently, hairpin probe biogate surface functionalized signal (MB@HP-Fe-MOF) cleaved activated Cas12a, leading release encapsulated electroactive molecule methylene blue, thereby generating strong signal. As result, this "AND" LC-CRISPR platform, requiring only single crRNA assembled achieves miRNA-155 miRNA-21 at concentrations low 3.2 fM. Moreover, allows easy adjustment according different targets, allowing be easily expanded analysis diagnosis other multibiomarkers. This approach demonstrates promising future applications intelligent diagnostic medicine.

Language: Английский

Logic Circuits for Intelligent Microcystin Monitoring Based on Aptamer Recognition and Toehold-Mediated Hairpin DNA Self-Assembly DOI Creative Commons
Jiafeng Pan, Ying Zeng, Yuyan Wang

et al.

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

Published: Jan. 17, 2025

A sensitive fluorescence biosensor was developed for microcystin-LR (MC-LR) detection using H1, H2, and H3 DNA probes as sensing elements. The aptamer in H1 can recognize the target. H2 labeled with FAM BHQ. MC-LR binding will activate self-assemblies through toehold-mediated strand displacement. In formed products (MC-LR/H1/nH2/nH3), BHQ be separated a high signal observed assay. is limit of 53 fM. We further constructed several logic circuits (AND-AND cascaded circuit, feedforward resource allocation circuit) MC-LR, MC-LA, MC-YR three inputs. numbers 0 1 are used to code input output signals. AND-AND cascade circuit produce only (111) combination. MC-LA signals, inhibit self-assembly execute negative operation. Through rational design probe hybridizations on four different magnetic beads (MBs), achieve an intelligent information. Our proposed not provide platform microcystin but also serve smart system sensing.

Language: Английский

Citations

1
A Primer-Regulated Rolling Circle Amplification (RCA) for Logic-Controlled Multiplexed Enzyme Analysis DOI
Shuiqin Chai, Weimin Sun, Xin Hou

et al.

ACS Applied Bio Materials, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 21, 2025

DNA-related enzymes are associated with various diseases and have been potential biomarkers for clinical diagnosis. Developing robust ultrasensitive methods is extremely favorable the detection of these biomarkers. To this purpose, a primer-regulated rolling circle amplification (RCA) strategy was ingeniously proposed. Briefly, RCA primer, which invalidated 3'-inverted dT (locked state) unable to initiate an reaction by phi29 DNA polymerase, embedded recognition substrate specific enzyme. In presence target, cleavage process enzyme prompted release regeneration 3'-OH (unlocked state), satisfying vital prerequisite RCA. By adopting programmable modular design, can be either single base sites or sequence different types enzymes. This also enables us conduct multiple conveniently, relying on logic-controlled manner including YES, OR, AND, AND-OR operations. Overall, proposed uniquely insightful provides universal tool analyses diverse

Language: Английский

Citations

1
One-step strand displacement-mediated nucleic acids signal-amplified analytical strategy based on superparamagnetism-functionalized DNA arrays DOI
Zhijun Gao, Qiumei Pu, Dongxia Li

et al.

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: unknown, P. 142596 - 142596

Published: March 1, 2025

Language: Английский

Citations

0
A rapid, specific, and simple-to-use biosensor for amplification-free determination of microRNA based on electrical potential-assisted and ternary hybridization DOI
Y. N. Liu, Yang Li,

Hao Gong

et al.

Microchimica Acta, Journal Year: 2025, Volume and Issue: 192(5)

Published: April 22, 2025

Language: Английский

Citations

0
Applications and development trend of biosensors in the detection and diagnosis of gastrointestinal tumors DOI Creative Commons

Cheng-Rong Zhang,

Lili Jin,

Feng Wang

et al.

BioMedical Engineering OnLine, Journal Year: 2025, Volume and Issue: 24(1)

Published: June 3, 2025

Gastrointestinal malignancies present a significant menace to the well-being of people across globe. Consequently, prompt and precise identification such tumors is utmost importance. Biosensor technology emerges as promising avenue in this regard, possessing considerable potential for detection diagnosis gastrointestinal tumors. This achieved through integration biometric components with physical chemical transducers, enabling rapid, sensitive, specific capabilities. In context colorectal cancer, notable advancements have been made nucleic acids, proteins, cells, exosomes. For instance, threshold miRNA has refined amole level, while methodologies diverse protein markers are under continuous improvement. addition, exosome techniques evolving steadily. Esophageal on other hand, exhibits its own characteristic enzyme cascade immune sensors, among others. Similarly, gastric there numerous innovative approaches miRNA, Looking ahead, performance biosensors tumor anticipated enhance further. The combination multiple joint expected gain prominence, realization miniaturization holds drive improvements treatment tumors, ultimately leading better patient outcomes more effective approach combating these life-threatening diseases.

Language: Английский

Citations

0
Acidic Extracellular pH-Activated Allosteric DNA Nanodevice for Fluorescence Imaging of APE1 Activity in Tumor Cells DOI
Hui He, Yuchen Wu, Mingjian Chen

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(45), P. 18079 - 18085

Published: Oct. 30, 2024

Allostery is a phenomenon where the binding of ligand at one allosteric site influences affinity for another an active site. Different from orthosteric regulation, it allows more precise control biomolecular activity and enhances stability molecules. Inspired by regulation natural molecules, we present Y-shaped DNA nanodevice, termed YssAP, that was pH-responsive functionalized with AS1411 aptamer accurate fluorescence imaging human apurinic/apyrimidinic endonuclease (APE1) in tumor cells. With rational design, YssAP could not be cut APE1, Cy5 proximity BHQ2, leading to suppressed signal emission. In contrast, since acidic pH acted as effector, underwent conformational change into activated probe (YdsAP) extracellular pH. After entering cell via specific recognition aptamer, overexpressed APE1 AP on YdsAP. moved far away resulting strong output. Compared direct construction substrate, nanodevices have effects, which can precisely adjusted changing switching state. We anticipate this strategy will applied screening inhibitors diagnosis.

Language: Английский

Citations

1
CRISPR/Cas12a-Powered Electrochemical Platform for Dual-miRNA Detection via an AND Logic Circuit DOI

Jiangbo Dong,

Changjun Hou, Liyuan Deng

et al.

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

Published: Dec. 24, 2024

The CRISPR/Cas technology shows great potential in molecular detection and diagnostics. However, it is still challenging to detect multiple targets simultaneously using the CRISPR-Cas system. Herein, we ingeniously leverage synergistic effect of two short single-stranded DNA activators construct a CRISPR/Cas12a-driven electrochemical sensing platform based on an AND logic circuit ("AND" LC-CRISPR) for simultaneous dual miRNAs. Specifically, exponential amplification reaction products triggered by dual-specific miRNAs are designed as binary inputs bind with Cas12a/crRNA, forming activating trans-cleavage ability CRISPR-Cas12a Subsequently, hairpin probe biogate surface functionalized signal (MB@HP-Fe-MOF) cleaved activated Cas12a, leading release encapsulated electroactive molecule methylene blue, thereby generating strong signal. As result, this "AND" LC-CRISPR platform, requiring only single crRNA assembled achieves miRNA-155 miRNA-21 at concentrations low 3.2 fM. Moreover, allows easy adjustment according different targets, allowing be easily expanded analysis diagnosis other multibiomarkers. This approach demonstrates promising future applications intelligent diagnostic medicine.

Language: Английский

Citations

1