scholarly journals Dynamic Aqueous Multiphase Reaction System for One-Pot CRISPR-Cas12a-Based Ultrasensitive and Quantitative Molecular Diagnosis

2020 ◽  
Vol 92 (12) ◽  
pp. 8561-8568 ◽  
Author(s):  
Kun Yin ◽  
Xiong Ding ◽  
Ziyue Li ◽  
Hui Zhao ◽  
Kumarasen Cooper ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
Reaction System ◽  
One Pot ◽  
Multiphase Reaction

scholarly journals Dynamic aqueous multiphase reaction system for simple, sensitive and quantitative one-pot CRISPR-Cas12a based molecular diagnosis

2020 ◽  
Author(s):  
Kun Yin ◽  
Xiong Ding ◽  
Ziyue Li ◽  
Hui Zhao ◽  
Kumarasen Cooper ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnosis ◽  
Molecular Diagnostics ◽  
Point Of Care ◽  
Sucrose Concentration ◽  
Reaction System ◽  
Fluorescent Detection ◽  
Quantitative Detection ◽  
One Pot ◽  
Multiphase Reaction

AbstractRecently, CRISPR-Cas technology has opened a new era of nucleic acid-based molecular diagnostics. However, current CRISPR-Cas-based nucleic acid biosensing has largely a lack of the quantitative detection ability and typically requires separate manual operations. Herein, we reported a dynamic aqueous multiphase reaction (DAMR) system for simple, sensitive and quantitative one-pot CRISPR-Cas12a based molecular diagnosis by taking advantage of density difference of sucrose concentration. In the DAMR system, recombinase polymerase amplification (RPA) and CRISPR-Cas12a derived fluorescent detection occurred in spatially separated but connected aqueous phases. Our DAMR system was utilized to quantitatively detect human papillomavirus (HPV) 16 and 18 DNAs with sensitivities of 10 and 100 copies within less than one hour. Multiplex detection of HPV16/18 in clinical human swab samples were successfully achieved in the DAMR system using 3D-printed microfluidic device. Furthermore, we demonstrated that target DNA in real human plasma samples can be directly amplified and detected in the DAMR system without complicated sample pre-treatment. As demonstrated, the DAMR system has shown great potential for development of next-generation point-of-care molecular diagnostics.

scholarly journals Zinc Chloride Catalyzed Amino Claisen Rearrangement of 1-N-Allylindolines: An Expedient Protocol for the Synthesis of Functionalized 7-Allylindolines

2019 ◽  
Vol 25 (1) ◽  
pp. 22-26 ◽  
Author(s):  
Seema Jain
Keyword(s):  
Zinc Chloride ◽  
Claisen Rearrangement ◽  
Reaction System ◽  
Biologically Active ◽  
Catalyst Loading ◽  
One Pot ◽  
Substitution Pattern ◽  
Claisen Rearrangements ◽  
Lower Temperature ◽  
Molecular Libraries

Abstract7-Allylindolines are valuable synthons for designing biologically active molecular libraries. Lewis acid catalyzed amino-Claisen rearrangement provides a one pot synthetic entry to these heteroarenes. In this context, Zinc chloride (ZnCl2)–N,N-dimethylformamide system efficiently catalyzed amino-Claisen rearrangements of 1-N-allylindolines to 7-allylindolines. The rearrangement is influenced by stereoelectronic effects of substituents present in 1-N-allylindolines. The substrates containing electron donating functionalities underwent rearrangement at lower temperature than substrates with electron withdrawing functional groups. The regioselectivity of the process is governed by the substitution pattern on allyl moiety in 1-N-allylindoline as well as ZnCl2 catalyst loading in the reaction system.

scholarly journals Tandem buildup of complexity of aromatic molecules through multiple successive electrophile generation in one pot, controlled by varying the reaction temperature

2016 ◽  
Vol 14 (5) ◽  
pp. 1680-1693 ◽  
Author(s):  
Akinari Sumita ◽  
Yuko Otani ◽  
Tomohiko Ohwada
Keyword(s):  
Reaction Temperature ◽  
Reaction System ◽  
Reaction Rates ◽  
Aromatic Molecule ◽  
One Pot ◽  
Aromatic Molecules

The unmasking reaction rates and the time of generation of highly reactive electrophiles can be controlled. This reaction system demonstrates the conceptual validity of one-pot build-up of a complex aromatic molecule from multiple starting components.

Novel boron chelate complexes from the reaction of salicylaldehydes, tertiary amines, and diphenylborinic or phenylboronic acid. Crystal and molecular structures of two new types of chelated organoborate salts

1997 ◽  
Vol 75 (9) ◽  
pp. 1203-1214 ◽  
Author(s):  
Wolfgang Kliegel ◽  
Jörg Metge ◽  
Steven J. Rettig ◽  
James Trotter
Keyword(s):  
Tertiary Amine ◽  
Phenylboronic Acid ◽  
Reaction System ◽  
Organoboron Compound ◽  
Ammonium Cation ◽  
Acid Anhydride ◽  
Molecular Structures ◽  
One Pot ◽  
Space Group ◽  
Three Component Reaction

The one-pot reaction of equimolar amounts of salicylaldehyde, diphenylborinic acid anhydride, and a tertiary amine in the presence of an alkanol (R′OH) led to the addition of R′OH to the aldehyde group and the formation of an O,O-acetal moiety within the chelate anion with the charge balanced by an ammonium cation arising from the tertiary amine. Exchanging the diphenylborinic acid in the three-component reaction system for phenylboronic acid did not give the analogous adduct or chelate but one additional mole equivalent of phenylboronic acid (anhydride) was incorporated, leading to a polycyclic anion containing pyroboronate and acetal boronate functions with an associated ammonium cation. Crystals of 1-methylpiperidinium 4-ethoxy-2,2-diphenyl-1,3-dioxa-2-borata-1,2,3,4-tetrahydronaphthalenate, 6d, are monoclinic, a = 9.522(1), b = 17.703(1), c = 14.974(1) Å, β = 98.275(10)°, Z = 4, space group P21/n, and those of 1-methylpiperidinium 6,10-epoxy-4-methoxy-6,8-diphenyl-5,7,9-trioxa-8-bora-6-borata-5,6,7,8,9,10-hexahydrobenzocyclooctenate, 10e, are monoclinic, a = 7.968(1), b = 19.3707(4), c = 15.9400(5) Å, β = 100.186(5)°, Z = 4, space group P21/n, The structures were solved by direct methods and refined by full-matrix least-squares procedures to R = 0.050 and 0.033 (Rw = 0.040 and 0.032) for 2815 and 3529 reflections with I ≥ 3σ(F2), respectively. Compounds 6d and 10e are both representative of new classes of organoboron chelates. Keywords: salicylaldehyde semiacetal diphenylboron chelate, salicylaldehyde bisboronate, organoboron compound, crystal structure.

scholarly journals Dual-Functional Peroxidase-Copper Phosphate Hybrid Nanoflowers for Sensitive Detection of Biological Thiols

2021 ◽  
Vol 23 (1) ◽  
pp. 366
Author(s):  
Xuan Ai Le ◽  
Thao Nguyen Le ◽  
Moon Il Kim
Keyword(s):  
Room Temperature ◽  
Reaction System ◽  
Clinical Diagnostics ◽  
Fluorescent Detection ◽  
One Pot ◽  
Catalytic Activities ◽  
Cellular Processes ◽  
Dual Functional ◽  
Copper Phosphate ◽  
Reliable Quantification

An effective strategy to detect biological thiols (biothiols), including glutathione (GSH), cysteine (Cys), and homocysteine (Hcy), holds significant incentive since they play vital roles in many cellular processes and are closely related to many diseases. Here, we demonstrated that hybrid nanoflowers composed of crystalline copper phosphate and horseradish peroxidase (HRP) served as a functional unit exhibiting dual catalytic activities of biothiol oxidase and HRP, yielding a cascade reaction system for a sensitive one-pot fluorescent detection of biothiols. The nanoflowers were synthesized through the anisotropic growth of copper phosphate petals coordinated with the amine/amide moieties of HRP, by simply incubating HRP and copper(II) sulfate for three days at room temperature. Copper phosphates within the nanoflowers oxidized target biothiols to generate H2O2, which activated the entrapped HRP to oxidize the employed Amplex UltraRed substrate to produce intense fluorescence. Using this strategy, biothiols were selectively and sensitively detected by monitoring the respective fluorescence intensity. This nanoflower-based strategy was also successfully employed for reliable quantification of biothiols present in human serum, demonstrating its great potential for clinical diagnostics.

Development of a novel one-pot reaction system utilizing a bifunctional Zr-based metal–organic framework

2014 ◽  
Vol 4 (3) ◽  
pp. 625 ◽  
Author(s):  
Takashi Toyao ◽  
Masakazu Saito ◽  
Yu Horiuchi ◽  
Masaya Matsuoka
Keyword(s):  
Metal Organic Framework ◽  
Reaction System ◽  
One Pot ◽  
Metal Organic ◽  
Organic Framework

Synthesis of Isoxazolines and Isoxazoles via Metal-Free Desulfitative Cyclization

Synthesis ◽  
2018 ◽  
Vol 50 (12) ◽  
pp. 2385-2393 ◽  
Author(s):  
Qiu Sun ◽  
Ling He ◽  
Jiaxin Cheng ◽  
Ze Yang ◽  
Yuansheng Li ◽  
...  
Keyword(s):  
Bond Formation ◽  
Reaction System ◽  
Hypervalent Iodine ◽  
Cascade Process ◽  
One Pot ◽  
Metal Free

A novel, one-pot reaction for the synthesis of isoxazolines and isoxazoles is developed via a cascade process under metal-free conditions. The approach involves the formation of intramolecular C–N and C–O bonds and intermolecular C–C bonds from aromatic alkenes or alkynes and N-hydroxysulfonamides using hypervalent iodine(VII) and iodine as the oxidant. Activation of C–H and C–C bonds/construction of C–O bonds/elimination of SO2/C–N bond formation is achieved in sequence­ in the reaction system.

scholarly journals An amino acid based system for CO2 capture and catalytic utilization to produce formates

2021 ◽  
Author(s):  
Duo Wei ◽  
Henrik Junge ◽  
Matthias Beller
Keyword(s):  
Amino Acid ◽  
Co2 Capture ◽  
Reaction System ◽  
Ambient Air ◽  
One Pot

A novel amino acid based reaction system for CO2 capture and utilization (CCU) to produce formates is presented applying a ruthenium-based catalyst. Noteworthy, CO2 can be captured from ambient air and converted to formates in one-pot (TON > 50 000).

scholarly journals A Thermostable Cas12b from Brevibacillus Leverages One-pot Detection of SARS-CoV-2 Variants of Concern

2021 ◽  
Author(s):  
Long T. Nguyen ◽  
Nicolas C. Macaluso ◽  
Brianna L.M. Pizzano ◽  
Melanie N. Cash ◽  
Jan Spacek ◽  
...  
Keyword(s):  
Thermal Instability ◽  
Reaction System ◽  
High Viral Load ◽  
Detection Methods ◽  
Clinical Samples ◽  
One Pot ◽  
Efficient Manner ◽  
Guide Rna ◽  
Design Requirements ◽  
The Cost

Current SARS-CoV-2 detection platforms lack the ability to differentiate among variants of concern (VOCs) in an efficient manner. CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) has the potential to transform diagnostics due to its programmability. However, many of the CRISPR-based detection methods are reliant on either a multi-step process involving amplification or elaborate guide RNA designs. A complete one-pot detection reaction using alternative Cas effector endonucleases has been proposed to overcome these challenges. Yet, current approaches using Alicyclobacillus acidiphilus Cas12b (AapCas12b) are limited by its thermal instability at optimum reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction temperatures. Herein, we demonstrate that a novel Cas12b from Brevibacillus sp. SYP-B805 (referred to as BrCas12b) has robust trans-cleavage activity at ideal RT-LAMP conditions. A competitive profiling study of BrCas12b against Cas12b homologs from other bacteria genera underscores the potential of BrCas12b in the development of new diagnostics. As a proof-of-concept, we incorporated BrCas12b into an RT-LAMP-mediated one-pot reaction system, coined CRISPR-SPADE (CRISPR Single Pot Assay for Detecting Emerging VOCs) to enable rapid, differential detection of SARS-CoV-2 VOCs, including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), and Delta (B.1.617.2) in 205 clinical samples. Notably, a BrCas12b detection signal was observed within 1-3 minutes of amplification, achieving an overall 98.1% specificity, 91.2% accuracy, and 88.1% sensitivity within 30 minutes. Significantly, for samples with high viral load (Ct value ≤ 30), 100% accuracy and sensitivity were attained. To facilitate dissemination and global implementation of the assay, we combined the lyophilized one-pot reagents with a portable multiplexing device capable of interpreting fluorescence signals at a fraction of the cost of a qPCR system. With relaxed design requirements, one-pot detection, and simple instrumentation, this assay has the capability to advance future diagnostics.

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