Enzyme-Free Crispr/Cas12a Microrna Assay by Amplified Single Particle Analysis DOI
Chengchao Zhang, Xin Zhao, Xiao Chen

et al.

Published: Jan. 1, 2024

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based detection platforms have increasingly been applied for microRNA (miRNA) detection, especially gained high sensitivity and wide application by combining enzyme-based nucleic acid pre-amplification strategies. Despite great success, the introduction of additional enzymes could result in negative interference between primer residues, higher costs, non-specific amplification which sometimes lead to false-positive outcomes or even deactivate system. To address these challenges, an enzyme-free amplified CRISPR/Cas12a miRNA assay was designed hybridization chain reaction (HCR) single particle analysis strategy. HCR utilizes self-assembly properties acids, avoids using any enzyme, enabling efficient mild conditions. Besides, inductively coupled plasma mass spectrometry (Sp-ICPMS) can provide a direct signal amplification, analyzing individual nanoparticles that retain number metal atoms. Herein, target miRNAs initiate circuit, be converted into double-strand DNAs (dsDNAs) with PAM regions recognized CRISPR/Cas The activated Cas12a would efficiently sabotage orthogonal nanoparticles-cross-linking system, generate various nano-aggregations different detectable Sp-ICPMS intensities. Eventually, fM level limit-of-detections (LODs) excellent selectivity were achieved. Serum-spiked recovery tests conducted promising results. our knowledge, this is first report HCR-CRISPR/Cas12a-based strategy detection.

Language: Английский

Demystifying EV heterogeneity: emerging microfluidic technologies for isolation and multiplexed profiling of extracellular vesicles DOI
Guihua Zhang, Xiaodan Huang, Sinong Liu

et al.

Lab on a Chip, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

The microfluidic-based technique that combines efficient isolation of EVs and multiple detection EV cargos like proteins, nucleic acids, glycans at bulk, single/single cell level to further demystify heterogeneity.

Language: Английский

Citations

0
The development of electrochemiluminescent probes: Mechanism and application DOI

Tongtong Li,

Yingying Su, Lichun Zhang

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113216 - 113216

Published: March 1, 2025

Language: Английский

Citations

0
Chiral sensing combined with nuclease activity assay to track Cas9 dynamics in solution: ROA and CPL study DOI Creative Commons
Monika Halat, Magdalena Klimek‐Chodacka, Agnieszka Domagała

et al.

Chemical Communications, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

We present a new non-destructive approach to detect CRISPR-Cas biomolecules and distinguish active from inactive forms using Raman optical activity (ROA) sensitive Eu( iii )-probes inducing circularly polarized luminescence (CPL).

Language: Английский

Citations

0
CRISPR/Cas12a dual-mode biosensor for Staphylococcus aureus detection via enzyme-free isothermal amplification DOI

Hongmin Gao,

Hehua Zhang,

Xue Qi

et al.

Talanta, Journal Year: 2024, Volume and Issue: 282, P. 127013 - 127013

Published: Oct. 10, 2024

Language: Английский

Citations

3
Recombinase Polymerase Amplification and Target-Triggered CRISPR/Cas12a Assay for Sensitive and Selective Hepatitis B Virus DNA Analysis Based on Lanthanide Tagging and Inductively Coupled Plasma Mass Spectrometric Detection DOI

Chenxi Zhao,

Lijie Du,

Jing Hu

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(37), P. 15059 - 15065

Published: Sept. 6, 2024

Herein, we report a target-triggered CRISPR/Cas12a assay by coupling lanthanide tagging and inductively coupled plasma mass spectrometry (ICP-MS) for highly sensitive elemental detection. Hepatitis B virus (HBV) DNA was chosen as model analyte, recombinase polymerase amplification (RPA) used target amplification. The double-stranded RPA amplicons containing 5' TTTG PAM sequence can be recognized Cas12a through specific CRISPR RNA, activating the

Language: Английский

Citations

2
CRISPR/Cas12a Assay for Amol Level microRNA by Combining Enzyme-free Amplification and Single Particle Analysis DOI
Chengchao Zhang, Xin Zhao, Xiao Chen

et al.

Chemical Communications, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 1, 2024

A new dual-amplification CRISPR-miRNA assay combining HCR and single-particle analysis mitigates PCR-related issues like high costs, non-specificity, primer interference.

Language: Английский

Citations

2
Nanomaterials as signal amplifiers in CRISPR/Cas biosensors: A path toward multiplex point-of-care diagnostics DOI
Fareeha Arshad,

Bong Jing Yee,

Koo Pey Ting

et al.

Microchemical Journal, Journal Year: 2024, Volume and Issue: 207, P. 111826 - 111826

Published: Oct. 2, 2024

Language: Английский

Citations

1
Enzyme-Free Crispr/Cas12a Microrna Assay by Amplified Single Particle Analysis DOI
Chengchao Zhang, Xin Zhao, Xiao Chen

et al.

Published: Jan. 1, 2024

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based detection platforms have increasingly been applied for microRNA (miRNA) detection, especially gained high sensitivity and wide application by combining enzyme-based nucleic acid pre-amplification strategies. Despite great success, the introduction of additional enzymes could result in negative interference between primer residues, higher costs, non-specific amplification which sometimes lead to false-positive outcomes or even deactivate system. To address these challenges, an enzyme-free amplified CRISPR/Cas12a miRNA assay was designed hybridization chain reaction (HCR) single particle analysis strategy. HCR utilizes self-assembly properties acids, avoids using any enzyme, enabling efficient mild conditions. Besides, inductively coupled plasma mass spectrometry (Sp-ICPMS) can provide a direct signal amplification, analyzing individual nanoparticles that retain number metal atoms. Herein, target miRNAs initiate circuit, be converted into double-strand DNAs (dsDNAs) with PAM regions recognized CRISPR/Cas The activated Cas12a would efficiently sabotage orthogonal nanoparticles-cross-linking system, generate various nano-aggregations different detectable Sp-ICPMS intensities. Eventually, fM level limit-of-detections (LODs) excellent selectivity were achieved. Serum-spiked recovery tests conducted promising results. our knowledge, this is first report HCR-CRISPR/Cas12a-based strategy detection.

Language: Английский

Citations

0