One-Pot Assay for Rapid Detection of Benzimidazole Resistance in Venturia carpophila by Combining RPA and CRISPR/Cas12a

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(3), P. 1381 - 1390

Published: Jan. 10, 2023

High resistance to benzimidazole fungicides in Venturia carpophila is caused by the point mutation E198K of β-tubulin (TUB2) gene. Traditional methods for detection fungicide are time-consuming, which routinely based on tedious operation, reliance expensive equipment, and specially trained people. Therefore, it important establish efficient field V. make suitable management strategies ensure food safety. Based recombinase polymerase amplification (RPA) combined with CRISPR/Cas12a, a rapid one-pot assay ORCas12a-BRVc (one-pot RPA-CRISPR/Cas12 platform) was established carpophila. The enabled adding components at bottom wall tube separately, solving problems aerosol contamination decreased sensitivity competing DNA substrates between Cas12a cleavage RPA amplification. could accomplish minimum 7.82 × 103 fg μL-1V. genomic 45 min 37 °C. Meanwhile, this showed excellent specificity due specific recognition ability Cas12a-crRNA complex. Further, we method that rapidly extract from within 2 achieve more simple fields. has advantages simplicity, rapidity, high sensitivity, specificity, ease operation without need precision instruments isolate culture pathogens. This first application platform combination CRISPR/Cas12a can be used monitoring resistant populations fields, providing guidance making peach scab.

Language: Английский

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CRISPR molecular detection techniques: Advances from single to multiple detection methods

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 166, P. 117198 - 117198

Published: July 24, 2023

Language: Английский

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CRISPR-powered quantitative keyword search engine in DNA data storage

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: March 15, 2024

Despite the growing interest of archiving information in synthetic DNA to confront data explosion, quantitatively querying stored is still a challenge. Herein, we present Search Enabled by Enzymatic Keyword Recognition (SEEKER), which utilizes CRISPR-Cas12a rapidly generate visible fluorescence when target corresponding keyword present. SEEKER achieves quantitative text searching since growth rate intensity proportional frequency. Compatible with SEEKER, develop non-collision grouping coding, reduces size dictionary and enables lossless compression without disrupting original order texts. Using four queries, correctly identify keywords 40 files background ~8000 irrelevant terms. Parallel can be performed on 3D-printed microfluidic chip. Overall, provides approach conducting parallel over complete content simple implementation rapid result generation.

Language: Английский

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Enhancing 3D DNA Walker-Induced CRISPR/Cas12a Technology for Highly Sensitive Detection of ExomicroRNA Associated with Osteoporosis

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(3), P. 1438 - 1446

Published: March 7, 2024

Exosomal microRNAs (exomiRNAs) have emerged as promising biomarkers for the early clinical diagnosis of osteoporosis. However, their limited abundance and short length in peripheral blood present significant challenges accurate detection exomiRNAs. Herein, we designed implemented an efficacious fluorescence-based biosensor highly sensitive exomiRNA associated with osteoporosis, leveraging enhancing 3D DNA walker-induced CRISPR/Cas12a technology. The engineered walker is capable efficiently transforming target into amplifying strands, thereby sensitivity developed biosensor. Concurrently, liberated strands serve activators to trigger Cas12a trans-cleavage activity, culminating a significantly amplified fluorescent signal exomiRNA-214. Under optimal conditions, devised technology demonstrated capacity detect exomiRNA-214 at concentrations low 20.42 fM, encompassing wide linear range extending from 50.0 fM 10.0 nM. Moreover, could accurately differentiate between healthy individuals osteoporosis patients via exomiRNA-214, which was agreement RT-qPCR results. As such, this biosensing offers promise valuable tool

Language: Английский

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Ultrasensitive protein and exosome analysis based on a rolling circle amplification assisted-CRISPR/Cas12a strategy

Talanta, Journal Year: 2024, Volume and Issue: 273, P. 125906 - 125906

Published: March 11, 2024

Language: Английский

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