Glutathione and biosensor technologies: enhancing plant resilience to environmental stressors

Physiological and Molecular Plant Pathology, Journal Year: 2025, Volume and Issue: unknown, P. 102570 - 102570

Published: Jan. 1, 2025

Language: Английский

---

Lighting up three-dimensional nanolantern for circular RNA imaging and precise gene therapy

Biosensors and Bioelectronics, Journal Year: 2025, Volume and Issue: 276, P. 117273 - 117273

Published: Feb. 15, 2025

Language: Английский

---

Logic-Measurer: A Multienzyme-Assisted Ultrasensitive Circuit for Logical Detection of Exosomal MicroRNAs

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

Published: March 19, 2025

The logic profiling of exosomal microRNAs (miRNAs) offers broad potential applications in the accurate diagnosis and staging cancer. However, logical detection low-abundance miRNAs complex clinical samples remains challenging. This study introduces a analysis system termed "Measurer" (a multi-enzyme-assisted ultrasensitive circuit) that versatile method for detecting multiple miRNAs. Logic-Measurer comprises three modules: stem-loop hairpin-enhanced CRISPR/Cas13a, polymerase-driven primer exchange reaction, an exonuclease III-mediated fluorescence output. efficient was switched by faster rate trans-cleavage activity Cas13a due to its improved affinity hairpin RNA structures. mechanistic model CRISPR/Cas13a confirmed molecular dynamics simulations. accurately detected miRNA-21 or miRNA-375 down 2.1 4.4 fM, with superior specificity, enabled situ as low 1.4 × 102 particles/mL exosomes via membrane fusion. In addition, this demonstrated 87.3 82.1% accuracy early breast cancer, respectively, among cohort 315 individuals. Subsequent subgroup further method's ability differentiate estrogen receptor-positive patients from healthy Therefore, valuable insights into development CRISPR/Cas-based enhanced diagnostic platform next generation technology based on enzyme circuits.

Language: Английский

---

Recent advances on CRISPR/Cas system-enabled portable detection devices for on-site agri-food safety assay

Trends in Food Science & Technology, Journal Year: 2022, Volume and Issue: 129, P. 364 - 387

Published: Oct. 4, 2022

Language: Английский

---

Low-Background CRISPR/Cas12a Sensors for Versatile Live-Cell Biosensing

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(42), P. 15725 - 15735

Published: Oct. 11, 2023

The trans-cleavage activity of CRISPR/Cas12a has been widely used in biosensing. However, many CRISPR/Cas12a-based biosensors, especially those that work "on-off-on" mode, usually suffer from high background and thus impossible intracellular application. Herein, this problem is efficiently overcome by elaborately designing the activator strand (AS) using "RESET" effect found our group. activation ability as-designed AS to can be easily inhibited, assuring a low for subsequent biosensing applications, which not only benefits detection sensitivity improvement biosensors but also promotes their applications live cells as well makes it possible design high-performance with greatly improved flexibility, achieving analysis wide range targets. As examples, different strategies such displacement, cleavage, aptamer-substrate interaction reactivate inhibited enzyme activity, several systems are developed sensitive specific targets, including nucleic acid (miR-21), biological small molecules (ATP), enzymes (hOGG1), giving limits 0.96 pM, 8.6 μM, 8.3 × 10-5 U/mL, respectively. Thanks background, these demonstrated accurate imaging biomolecules cells. Moreover, we demonstrate sensing combined lateral flow assay (LFA), holding great potential point-of-care testing, poorly equipped or nonlaboratory environments.

Language: Английский

---

CRISPR/Cas-Assisted Nanoneedle Sensor for Adenosine Triphosphate Detection in Living Cells

ACS Applied Materials & Interfaces, Journal Year: 2023, Volume and Issue: 15(43), P. 49964 - 49973

Published: Sept. 28, 2023

The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) (CRISPR/Cas) systems have recently emerged as powerful molecular biosensing tools based on their collateral cleavage activity due to simplicity, sensitivity, specificity, and broad applicability. However, the direct application of for in situ intracellular detection is still challenging. Here, we debut a CRISPR/Cas-assisted nanoneedle sensor (nanoCRISPR) adenosine triphosphate (ATP), which avoids challenges associated with by introducing two-step process target recognition, followed extracellular transduction detection. ATP recognition occurs first presenting cell cytosol an aptamer-locked Cas12a activator conjugated nanoneedles; event unlocks immobilized nanoneedles. nanoneedles are then removed from cells exposed Cas12a/crRNA complex, where triggers ssDNA fluorophore-quencher pair, generating detectable fluorescence signal. NanoCRISPR has limit 246 nM dynamic range 1.56 50 μM. Importantly, nanoCRISPR can detect 30 min live without impacting viability. We anticipate that approach will contribute broadening biomedical applications CRISPR/Cas sensors diverse molecules living systems.

Language: Английский

---

Ultrasensitive miRNA Detection Based on Magnetic Upconversion Nanoparticle Enhancement and CRISPR/Cas13a-Driven Signal Amplification

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(48), P. 17708 - 17715

Published: Nov. 24, 2023

MicroRNAs (miRNAs), a class of small molecules with important regulatory functions, have been widely used in the field biosensing as biomarkers for early diagnosis various diseases. Therefore, it is crucial to develop an miRNA detection platform high sensitivity and specificity. Here, we designed CRISPR/Cas13-based enzymatic cyclic amplification system regarded magnetic upconversion nanoparticles (MUCNPs) biosensor outputting signal highly sensitive high-fidelity miRNAs. MUCNPs were composed UCNPs (fluorescence donors) Fe3O4@AuNPs acceptors) through double-stranded DNA hybrid coupling. The target acted activator, which could activate trans-cleavage activity Cas13a well-designed Trigger containing two uracil ribonucleotides (rU) its loop trigger strand displacement reaction generate large amount single-stranded DNA, resulting release from MUCNPs. Benefiting fidelity selectivity CRISPR/Cas13a, great effect triggered cycle amplification, high-intensity luminescent MUCNPs, this method possessed capability specificity even complex environment 10% fetal bovine serum (FBS) sample. Meanwhile, limit be low 83.2 fM. In addition, effectively reduced photobleaching maintained stability, was expected achieve efficient detection.

Language: Английский

---

DNA Gate-Based CRISPR-Cas Exponential Amplification System for Ultrasensitive Small Extracellular Vesicle Detection to Enhance Breast Cancer Diagnosis

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(3), P. 1328 - 1335

Published: Jan. 8, 2024

Tumor-derived small extracellular vesicles (tEVs) as potential biomarkers possess abundant surface proteins closely related to parent cells, which are crucial for noninvasive cancer diagnosis. However, tEVs exhibit phenotype heterogeneity and low abundance, posing a significant challenge multiplex detection with high sensitivity. Herein, we developed DNA gate-based exponential amplification CRISPR-Cas (DGEAC) system accurate ultrasensitive of tEVs, can greatly improve the accuracy breast (BC) Based on coexpression CD63 vascular endothelial growth factor (VEGF) BC-derived dual-aptamer-based AND gate fluorescent probe by proximity hybridization. By integrating target recognition trans-cleavage activity Cas12a, an autocatalysis-driven circuit was VEGF could avoid false negative signals from single protein or other interfering proteins. We achieved highly sensitive over linear range 1.75 × 103 3.5 108 particles/mL limit 1.02 particles/mL. Furthermore, DGEAC distinguish derived different BC cell lines, including MDA-MB-231, MCF-7, SKBR3, MCF-10A. Compared (AUC 90.0%), effectively differentiates in stages 98.3%).

Language: Английский

---

DNA Robots for CRISPR/Cas12a Activity Management and Universal Platforms for Biosensing

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(6), P. 2620 - 2627

Published: Jan. 13, 2024

The CRISPR/Cas12a system is a revolutionary genome editing technique that widely employed in biosensing and molecular diagnostics. However, there are few reports on precisely managing the trans-cleavage activity of Cas12a by simple modification since traditional methods to manage often require difficult rigorous regulation core components. Hence, we developed novel regulatory mechanism, named DNA Robots for Enzyme Activity Management (DREAM), introducing two robots, apurinic/apyrimidinic site (AP site) or nick target activator. First, revealed mechanism how DREAM strategy regulated through different binding affinities. Second, was found improve selectivity identifying base mismatch. Third, modular biosensor excision repair enzymes based utilizing diversified generation ways multi-signal output platform such as fluorescence, colorimetry, visual lateral flow strip constructed. Furthermore, extended logic sensing circuits overcome barrier could not detect simultaneously single tube. Overall, only provided new prospects programmable systems but also enabled portable, specific, humanized detection with great potential

Language: Английский

---

Regulating the trans-Cleavage Activity of CRISPR/Cas12a by Using an Elongation-Caged Single-Stranded DNA Activator and the Biosensing Applications

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(32), P. 12169 - 12176

Published: Aug. 2, 2023

The CRISPR/Cas12a system exhibits extraordinary capability in the field of biosensing and molecular diagnosis due to its trans-cleavage ability. However, it is still desirable for precise control programmable regulation Cas12a activity promote in-depth studies application expansion Cas12a-based sensing platforms. In this work, we have developed a new robust mechanism by endowing activator with function caging crRNA ingeniously. Specifically, constructed an integrated elongation-caged (EL-activator) extending ssDNA on 3′-end. We found that appending only about 8 nt complementary repeat region enough cage spacer/repeat region, thus effectively inhibiting activity. inner inhibition was further uncovered after thorough investigation, demonstrating EL-activator works impeding conformation required recognition destroying affinity Cas12a. By switching elongated moiety using target biomarkers, blocked can be rapidly recovered. Finally, versatile platform established based mechanism, expanding conventional directly recognizes DNA direct detection enzymes RNA biomarkers. This work has enriched toolbox expanded applications.

Language: Английский

---