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TALANTA · 2021 · IF 6.057

A strategy combining 3D-DNA Walker and CRISPR-Cas12a trans-cleavage activity applied to MXene based electrochemiluminescent sensor for SARS-CoV-2 RdRp gene detection

Kai Zhang,Zhenqiang Fan,Yue Huang,Yuedi Ding,Minhao Xie

Early diagnosis and timely management of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) are the keys to preventing the spread of the epidemic and controlling new infection clues. Therefore, strengthening the surveillance of the epidemic and timely screening and confirming SARS-CoV-2 infection is the primary task. In this work, we first proposed the idea of activating CRISPR-Cas12a activity using double-stranded DNA amplified by a three-dimensional (3D) DNA walker. We applied it to the design of an electrochemiluminescent (ECL) biosensor to detect the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene. We first activated the cleavage activity of CRISPR-Cas12a by amplifying the target DNA into a segment of double-stranded DNA through the amplification effect of a 3D DNA walker. At the same time, we designed an MXene based ECL material: [email protected] 3 C 2 @AuNPs, and constructed an ECL biosensor to detect the RdRp gene based on this ECL material as a framework. Activated CRISPR-Cas12a cleaves the single-stranded DNA on the surface of this sensor and causes the ferrocene modified at one end of the DNA to move away from the electrode surface, increasing the ECL signal. The extent of the change in electrochemiluminescence reflects the concentration of the gene to be measured. Using this system, we detected the SARS-CoV-2 RdRp gene with a detection limit of 12.8 aM. This strategy contributes to the rapid and convenient detection of SARS-CoV-2-associated nucleic acids and promotes the clinical application of ECL biosensors based on CRISPR-Cas12a and novel composite materials.

研究领域: 生物技术 · 纳米技术 · 分子生物学 · 病毒学 · 分析化学
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MICROCHEMICAL JOURNAL · 2023 · IF 4.8

An electrochemiluminescence sensing platform combining entropy-driven amplification strategy and inverted DNA tetrahedral nanomachines for detecting SARS-CoV-2 specific gene

Jiaying Wang,Bin Li,Bo Yao,Yi Zhao,Kai Zhang

In this work, we propose a Ti 3 C 2 T x -based electrochemiluminescence (ECL) sensing platform, combined with a programmable entropy-driven cycling strategy and inverted DNA tetrahedral nanomachines to detect the SARS-CoV-2 RdRp gene. The use of Ti 3 C 2 T x in the ECL sensing platform provides unique advantages that improve the assay's sensitivity. Firstly, the SARS-CoV-2 RdRp gene triggers an entropy-driven cyclic amplification reaction to obtain a large number of Output strands. Subsequently, the Output strand reacts with an inverted DNA tetrahedral swing arm strand to start the self-driven process. Finally, the ferrocene (Fc)-labeled hairpin DNA strand of the inverted DNA tetrahedron is severed to achieve the signal change. Under optimal conditions, the ECL platform integrated with entropy-driven cycle amplification reaction and inverted DNA tetrahedral nanomachines obtained a wide range of 100 aM to 10 nM and a limit of detection (LOD) of 48.3 aM for the SARS-CoV-2 RdRp gene. Moreover, the recovery rates of 97% to 103.3% and 98.34% to 103% were obtained in real environments and human pharyngeal swabs, respectively. This ECL sensing platform, which combines an entropy-driven cycle amplification reaction and inverted DNA tetrahedral nanomachines, opens up new opportunities for the detection of other substances.

研究领域: 生物化学 · 纳米技术 · 分析化学 · 分子生物学 · 病毒学
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VETERINARNI MEDICINA · 2023 · IF 0.7

Ribavirin inhibits peste des petits ruminants virus proliferation in vitro

Weifeng Zhang,Hualong Deng,Yanfen Liu,Shaohong Chen,You Liu,Yuntao Zhao

Peste des petits ruminants virus (PPRV), a member of the family Paramyxoviridae, belongs to the genus Morbillivirus. It causes devastating viral diseases in small ruminants and has been rapidly spreading over various regions in Africa, the Middle East, and Asia. Although vaccination is thought to be an effective management strategy against PPR infections, the heat sensitivity of PPRV vaccines severely restricts their use in regions with hot climates. In this research, we studied the antiviral activities of ribavirin and aimed to understand the potential mechanisms of action of ribavirin in the African green monkey kidney cells (Vero cells). In brief, the adsorption, intrusion, replication, and release of PPRV, as well as the mRNA expression level of RNA-dependent RNA polymerase ( RdRp ), were significantly inhibited in the ribavirin-treated Vero cells compared to those in the PPRV-infected cells that were not treated with ribavirin. Additionally, ribavirin has potential as an antiviral drug against PPRV, and its antiviral activity is mediated by the Janus kinase signal transducer and activator of transcription (JAK/STAT) and PI3K/AKT pathways.

CAS / MDL: 36791-04-5
研究领域: 兽医学 · 病毒学 · 分子生物学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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CHEMICAL ENGINEERING JOURNAL · 2021 · IF 13.273

Rational engineering the DNA tetrahedrons of dual wavelength ratiometric electrochemiluminescence biosensor for high efficient detection of SARS-CoV-2 RdRp gene by using entropy-driven and bipedal DNA walker amplification strategy

Zhenqiang Fan,Bo Yao,Yuedi Ding,Dong Xu,Jianfeng Zhao,Kai Zhang

Fast and effective detection of epidemics is the key to preventing the spread of diseases. In this work, we constructed a dual-wavelength ratiometric electrochemiluminescence (ECL) biosensor based on entropy-driven and bipedal DNA walker cycle amplification strategies for detection of the RNA-dependent RNA polymerase (RdRp) gene of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The entropy-driven cyclic amplification reaction was started by the SARS-CoV-2 RdRp gene to generate a bandage. The bandage could combine with two other single-stranded S1 and S2 to form a bipedal DNA walker to create the following cycle reaction. After the bipedal DNA walker completed the walking process, the hairpin structures at the top of the DNA tetrahedrons (TDNAs) were removed. Subsequently, the [email protected] 3 C 2 @AuNPs-S7 probes were used to combine with the excised hairpin part of TDNAs on the surface of Au-g-C 3 N 4 , and the signal change was realized employing electrochemiluminescence resonance energy transfer (ECL-RET). By combining entropy-driven and DNA walker cycle amplification strategy, the ratiometric ECL biosensor exhibited a limit of detection (LOD) as low as 7.8 aM for the SARS-CoV-2 RdRp gene. As a result, detecting the SARS-CoV-2 RdRp gene in human serum still possessed high recovery so that the dual-wavelength ratiometer biosensor could be used in early clinical diagnosis.

研究领域: 生物技术 · 分析化学 · 分子生物学 · 纳米技术 · 病毒学
✓ 已验证 DOI PubMed 高影响因子
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INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH · 2020 · IF 3.573

Novel PVDF-g-NMA Copolymer for Fabricating the Hydrophilic Ultrafiltration Membrane with Good Antifouling Property

Yujia Tong,Lukuan Huang,Chengjiang Zuo,Weixing Li,Weihong Xing

Improving the hydrophilicity of the poly(vinylidene fluoride) (PVDF) ultrafiltration membrane can change its adsorption fouling characteristics. Here, a novel amphiphilic material of PVDF grafted with N-methylolacrylamide (PVDF-g-NMA) was developed via photoinduced Cu(II)-mediated reversible deactivation radical polymerization (RDRP). The PVDF-g-NMA ultrafiltration membrane was prepared by the nonsolvent-induced phase-separation method. The PVDF-g-NMA copolymer was characterized by 1H NMR and Fourier transform infrared spectroscopy. The performance of the PVDF-g-NMA membrane was evaluated by determining the permeation flux, contact angle, roughness, and antifouling tests. The roughness data show that a low content of N-methylolacrylamide can effectively reduce the surface roughness of membranes by 50%, resulting in greater antifouling ability. The prepared ultrafiltration membrane containing 15 wt % PVDF-g-NMA exhibited the best hydrophilicity with an average pure water flux of up to 272.1 L·m–2·h–1, higher than that of the pure PVDF membrane (45.4 L·m–2·h–1). The contact angle of the PVDF-g-NMA ultrafiltration membrane decreased from 85.5 to 67.4°. The bovine serum albumin rejection rate of the PVDF-g-NMA membrane significantly increased from 85.7 to 92.6%, and the pure water flux recovery rate increased from 79.0 to 88.5%. The PVDF-g-NMA ultrafiltration membrane had excellent hydrophilicity and antifouling properties, which would be promising for wastewater treatment.

CAS / MDL: 7447-39-4 924-42-5
研究领域: 材料科学 · 环境工程 · 化学工程
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JOURNAL OF POLYMER RESEARCH · 2023 ·Vol. 30 ·期 9 ·1-8 · IF 2.8

Preparation of PDMS STEM gels through oxygen tolerance nitroxide mediated polymerization

Qiao Xiaoguang,Dong Xin,shi Ge,He Yanjie,Pang Xinchang

The postsynthesis polymeric modifications by grafting from the active sites were the primary focus of the structurally tailored and engineered macromolecular (STEM) gels. Therefore, Reversible-deactivation radical polymerization (RDRP) techniques have been employed to more precisely tailor and tune STEM gels. In contrast to atom transfer radical polymerization (ATRP) and reversible addition-fragmentation chain transfer polymerization (RAFT), nitroxide-mediated polymerization (NMP) is known to have the advantages of simple components and pure products. Here we report the preparation of polydimethylsiloxane (PDMS) composite gel networks through oxygen tolerance NMP process, with tert-Butyl acrylate (tBA) and fluorinated monomer 2-(Perfluorooctyl)ethyl methacrylate (FMA) as functional comonomers. Compared with conventional radical polymerization (FRP) method, composite gels prepared through NMP method showed higher swelling rate and elongation at break, which proved that the network structure formed by NMP was more uniform. In addition to better thermal stability and toughness, the dormant polymer chain could be reactivated and lead to the “living” post-modification of PDMS gel matrix.

研究领域: 材料科学 · 高分子化学 · 材料工程
✓ 已验证 产品关联 DOI 高影响因子
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Environmental Science-Nano · 2021 ·Vol. 9 ·期 1 ·162-172 · IF 8.131

Exploring the entropy-driven amplification reaction and trans-cleavage activity of CRISPR-Cas12a for the development of an electrochemiluminescence biosensor for the detection of the SARS-CoV-2 RdRp gene in real samples and environmental surveillance

Kai Zhang,Zhenqiang Fan,Yuedi Ding,Sha Zhu,Minhao Xie,Nan Hao

In this paper, we present the first idea of using a DNA triple helix structure to inhibit CRISPR-Cas12a activity and apply it to the design of an electrochemiluminescent biosensor for the detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA-dependent RNA polymerase (RdRp) gene in real samples and environmental surveillance. We employed a segment from the RdRp gene of SARS-CoV-2 by an entropy-driven reaction, which was paired with double-stranded DNA that can activate CRISPR-Cas12a activity by Hoogsteen pairing to form triple-stranded DNA, thereby inhibiting the binding interaction of the double-stranded DNA with CRISPR-Cas12a, which in turn inhibits the trans cleavage activity of CRISPR-Cas12a. The inhibited CRISPR-Cas12a is unable to cut the nucleic acid modified on the electrode surface, resulting in the inability of the ferrocene (Fc) modified on the other end of the nucleic acid to move away from the electrode surface, and thus failing to cause electrochemiluminescence changes in GOAu–Ru modified on the electrode surface. The extent of the electrogenic chemiluminescence change can reflect the concentration of the gene to be tested. Using this system, we achieved the detection of the SARS-CoV-2 RdRp gene with a detection limit of 32.80 aM.

研究领域: 分子生物学 · 生物技术 · 分析化学 · 病毒学
✓ 已验证 DOI 高影响因子
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Polymer Chemistry · 2020 ·Vol. 11 ·期 47 ·7497-7505 · IF 5.342

Photocontrolled iodine-mediated reversible-deactivation radical polymerization with a semifluorinated alternating copolymer as the macroinitiator

Jiannan Cheng,Kai Tu,Enjie He,Jinying Wang,Lifen Zhang,Zhenping Cheng,Xiulin Zhu

A new photocontrolled iodine-mediated reversible-deactivation radical polymerization (RDRP) strategy to facilely prepare main-chain-type semifluorinated alternating block copolymers was successfully established under irradiation with blue LED light at room temperature, by using a semifluorinated alternating copolymer ((AB)n, where A represents α,ω-diiodoperfluoroalkane and B represents α,ω-unconjugated diene) as a macroinitiator and chain-transfer agent, and methacrylates, acrylates and styrene as the monomers. The end group of (AB)n terminated with perfluoroalkyl iodide –(CF2)n–I can be activated by Ru(bpy)3Cl2/AsAc-Na reducing quenching catalytic cycle which generates initial carbon centered radicals (–(CF2)n˙) to initiate the polymerization under blue LED light irradiation. The polymerization can reach a reversible equilibrium through effective oxygen–iodine halogen bonding in acetone and undergo a photocontrolled iodine-mediated RDRP process. In addition, the “living” features of this photocontrolled iodine-mediated RDRP system were confirmed by polymerization kinetics and controlled “on–off” light switching cycle regulation. What's more, different types of fluoropolymers can be introduced on the copolymer chains due to the powerful designability of (AB)n. Therefore, this strategy can provide a facile and highly efficient approach to the precision synthesis of main-chain-type semifluorinated alternating di/triblock polymers under visible light irradiation.

研究领域: 高分子化学 · 材料科学 · 光化学 · 有机化学
✓ 已验证 DOI 高影响因子
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