Self-Sustainable Catalytic Nucleic Acid Circuit Powered Hybrid Crispr-Cas Systems for Point-of-Care Diagnosis of Circadian Clock Gene from Cell Lysates DOI
Zhiyuan Feng, Ran Liu,

Enming Yang

et al.

Published: Jan. 1, 2024

The CRISPR-Cas13 systems have opened a new avenue for RNA detection due to their exceptional programmable collateral activity against ssRNA. However, most high-performance Cas13-based sensors still suffer from some drawbacks because of the tradeoffs between readout device complexity and analytical sensitivity, which can limit viability at point-of-care (POC). To overcome these shortcomings, we herein report novel target-responsive POC platform translating into glucose test. Specifically, this combines advantages three techniques: hybrid Cas13a/Cas12a system both recognition with single-base resolution cascade enzymatic amplification, self-sustainable catalytic nucleic acid circuit (abbreviated as SCC) embedded uracil-base poly-T bulges Cas13a- Cas12a-mediated cleavage, respectively, portable meter (PGM) simple signal readout. incorporation an engineered ssDNA–invertase conjugate in SCC led RNA-to-glucose transduction through hydrolysis sucrose glucose. By targeting 20-nt conserved region BMAL1 mRNA (mBMAL1), key circadian clock gene, demonstrate that serve excellent CRISPR reporter, enabling direct unamplified mBMAL1 low 0.864 fM concurrently good discrimination ability mismatch. More importantly, integrated was successfully applied diagnosis different cell lysates, results strongly correlate RT-PCR. Given its wide adaptability, anticipate such CRISPR-SCC be easily modified quantify other biomarkers associated rhythm.

Language: Английский

Self-sustainable Catalytic Nucleic Acid Circuit Powered Hybrid CRISPR-Cas Systems for Point-of-Care Diagnosis of Circadian Clock Gene from Cell Lysates DOI
Zhiyuan Feng, Ran Liu,

Enming Yang

et al.

Journal of Analysis and Testing, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 20, 2024

Language: Английский

Citations

2
CRISPR for companion diagnostics in low-resource settings DOI Creative Commons
Xu Qian, Qiang Xu, Christopher J. Lyon

et al.

Lab on a Chip, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 1, 2024

New point-of-care tests (POCTs), which are especially useful in low-resource settings, needed to expand screening capacity for diseases that cause significant mortality: tuberculosis, multiple cancers, and emerging infectious diseases. Recently, clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostic (CRISPR-Dx) assays have emerged as powerful versatile alternatives traditional nucleic acid tests, revealing a strong potential meet this need new POCTs. In review, we discuss CRISPR-Dx assay techniques been or could be applied develop POCTs, including sample processing, target amplification, multiplex design, signal readout. This review also describes current applications POCTs disease diagnosis includes future opportunities challenges such tests. These advance beyond initial development efforts broadly criteria use settings.

Language: Английский

Citations

1
Quencher-free fluorescent assays by controlled DNA partitioning in the aqueous two-phase system with crowding-enhanced kinetics DOI

Haoyue Lv,

Xili Duan, Zhaoyu Han

et al.

Biosensors and Bioelectronics, Journal Year: 2023, Volume and Issue: 246, P. 115864 - 115864

Published: Nov. 25, 2023

Language: Английский

Citations

2
Self-Sustainable Catalytic Nucleic Acid Circuit Powered Hybrid Crispr-Cas Systems for Point-of-Care Diagnosis of Circadian Clock Gene from Cell Lysates DOI
Zhiyuan Feng, Ran Liu,

Enming Yang

et al.

Published: Jan. 1, 2024

The CRISPR-Cas13 systems have opened a new avenue for RNA detection due to their exceptional programmable collateral activity against ssRNA. However, most high-performance Cas13-based sensors still suffer from some drawbacks because of the tradeoffs between readout device complexity and analytical sensitivity, which can limit viability at point-of-care (POC). To overcome these shortcomings, we herein report novel target-responsive POC platform translating into glucose test. Specifically, this combines advantages three techniques: hybrid Cas13a/Cas12a system both recognition with single-base resolution cascade enzymatic amplification, self-sustainable catalytic nucleic acid circuit (abbreviated as SCC) embedded uracil-base poly-T bulges Cas13a- Cas12a-mediated cleavage, respectively, portable meter (PGM) simple signal readout. incorporation an engineered ssDNA–invertase conjugate in SCC led RNA-to-glucose transduction through hydrolysis sucrose glucose. By targeting 20-nt conserved region BMAL1 mRNA (mBMAL1), key circadian clock gene, demonstrate that serve excellent CRISPR reporter, enabling direct unamplified mBMAL1 low 0.864 fM concurrently good discrimination ability mismatch. More importantly, integrated was successfully applied diagnosis different cell lysates, results strongly correlate RT-PCR. Given its wide adaptability, anticipate such CRISPR-SCC be easily modified quantify other biomarkers associated rhythm.

Language: Английский

Citations

0