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Nanomechanical assay for ultrasensitive and rapid detection of SARS-CoV-2 based on peptide nucleic acid

Nano Research 2023年第1期16卷 1183-1195页

作者： Yu Wang Tianhao Yan Kainan Mei Depeng Rao Wenjie Wu Ye Chen Yongpei Peng Jianye Wang Shangquan Wu Qingchuan Zhang CAS Key Laboratory of Mechanical Behavior and Design of Material Department of Modern MechanicsUniversity of Science and Technology of ChinaHefei 230027China Reproductive Medicine Center Department of Obstetrics and GynecologyFirst Affiliated Hospital of Anhui Medical UniversityHefei 230022China

The massive global spread of the COVID-19 pandemic makes the development of more effective and easily popularized assays ***,we developed an ultrasensitive nanomechanical method based on microcantilever array and peptide nucleic acid(PNA)for the detection of severe acute respiratory syndrome-coronavirus-2(SARS-CoV-2)*** method has an extremely low detection limit of 0.1 fM(105 copies/mL)for N-gene specific sequence(20 bp).Interestingly,it was further found that the detection limit of N gene(pharyngeal swab sample)was even lower,reaching 50 copies/*** large size of the N gene dramatically enhances the sensitivity of the nanomechanical sensor by up to three orders of *** detection limit of this amplification-free assay method is an order of magnitude lower than RT-PCR(500 copies/mL)that requires *** non-specific signal in the assay is eliminated by the in-situ comparison of the array,reducing the false-positive misdiagnosis *** method is amplification-free and label-free,allowing for accurate diagnosis within 1 *** strong specificity and ultrasensitivity allow single base mutations in viruses to be distinguished even at very low ***,the method remains sensitive to fM magnitude lung cancer marker(miRNA-155).Therefore,this ultrasensitive,amplification-free and inexpensive assay is expected to be used for the early diagnosis of COVID-19 patients and to be extended as a broad detection tool.

关键词： SARS-CoV-2 nucleic acid assay nanomechanical sensoring peptide nucleic acid amplification-free

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Synthesis and photo activity of phenylazonaphthalene peptide nucleic acid monomers

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Chinese Chemical Letters 2008年第7期19卷 807-810页

作者： Jin Du Li Miao Chen Sheng Liu Hao Bo Zhang Zhi Lu Liu Lanzhou Institute of Chemical Physics Chinese Academy of Sciences Lanzhou 730000 PR China Graduate School of Chinese Academy of Sciences Beijing 100039 PR China

Phenylazonaphthalene peptide nucleic acid （PNA） monomers were successfully synthesized, and their photoisomerization was examined. The new PNA monomers showed reversible trans-cis isomerization with UVand visible light irradiation, which might be the foundation of photo-regulating the hybridization between PNA containing phenylazonaphthalene unit and DNA. Simultaneously, the fluorescence of the new PNA monomers might make them especially useful as structural probes.

关键词： peptide nucleic acid Phenylazonaphthalene Photoisomerization UV-vis absorption spectrometry Fluorescence spectrometry

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peptide nucleic acid:a new artificial biomacromolecular with great potential applications in molecular biology and biom edicine

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Chinese Journal of Biomedical Engineering(English Edition) 2001年第4期10卷 165-168页

作者： WANG Jin-ke LU Zu-hong HE Nong-Yue Laboratory for Molecular and Biomolecular Electronics Southeast University Nanjing 210096 China College of Life Sciences Nanjing Normal University Nanjing 210097China Laboratory for Molecular and Biomolecular Electronics Southeast University Nanjing 210096 China

peptide nucleic acid (PNA) is a DNA mimic that was originally developed by Peter E Nielsen in 1991 as a reagent for sequence-specific recognition of double stranded (ds) DNA by a conventional triple helix type principle.

关键词： PNA acid peptide nucleic acid

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Electrochemical investigation of DNA hybridization with ferrocene-labelled peptide nucleic acid on gold electrodes

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Journal of Chongqing University 2010年第2期9卷 100-104页

作者：张浩波杨晰 College of Science Gansu Agricultural University

An electrochemical DNA sensor based on ferrocene-labelled peptide nucleic acid （PNA-Fc） was prepared. The hybridization between PNA-Fc and DNA immobilized on a gold electrode was examined by cyclic voltammetry （CV） and differential pulse voltammetry （DPV）. PNA-Fc shows a good electrochemically activity and has a redox potential of 170 mV verus Ag/AgCl electrode after hybridization, representing the characteristic of ferrocene/ferrocenium （Fc/Fc＋） transformation. The results illustrate that PNA-Fe can be used as an effective electrochemical DNA probe sensor.

关键词： peptide nucleic acid ferrocene electrochemical sensor

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Effects of peptide nucleic acids against Ki-67 Gene on the Proliferation and Apoptosis of Human Renal Carcinoma Cell Line

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第6期26卷 703-705页

作者：赖海标郑骏年吴松孙晓青陈家存 Department of Urology Affiliated Zhongshan Hospital of Guangzhou Traditional Chinese Medicine University Laboratory of Urology Affiliated Hospital of Xuzhou Medical College

To investigate the effects of anti-sense peptide nucleic acids （PNAs） targeting Ki-67 gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations （1.0 μmol/L, 2.0 μmol/L, 10.0 μmol/L）. The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and ^3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide （ASODNs） targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs （16.9±0.7） was significantly inhibited as compared with that of the control groups （28.6±0.4）（P〈0.01）. The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs （42.1 ±2.2） was significantly reduced when compared with that of the control groups （83.6± 1.4）（P〈0.01）. Proliferation of 786-0 cells treated with anti-sense PNAs （20.7 ± 1.5） was significantly inhibited as compared with that of the control groups （58.6± 1.4）（P〈0.01）. The apoptosis rate of 786-0 cells treated with anti-sense PNAs （28.7 ± 2.3） was significantly increased higher compared with that of the control groups （13.8 ±1.0）（P〈0.01）. From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.

关键词： peptide nucleic acid antisense oligonucleotide Ki-67 gene renal cell carcinoma proliferation apoptosis

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Essential Gene(s) Targeted by peptide nucleic acids Kills <i>Mycobacterium smegmatis</i>in Culture and in Infected Macrophages

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Advances in Infectious Diseases 2021年第2期11卷 156-164页

作者： Md. Ariful Islam Mst. Minara Khatun Nammalwar Sriranganathan Stephen M. Boyle Center for One Health Research Virginia-Maryland College of Veterinary Medicine Virginia Tech Blacksburg VA USA Department of Microbiology & Hygiene Bangladesh Agricultural University Mymensingh Bangladesh

Background: Antisense peptide nucleic acids (PNAs) exhibit growth inhibitory effects on bacteria by inhibiting the expression of essential genes and could be promising therapeutic agents for treating bacterial infections. A study was carried out to determine the efficacy of several antisense PNAs in inhibiting extracellular and intracellular growth of Mycobacterium smegmatis. Methods: Six PNAs obtained from a commercial supplier were tested to evaluate the inhibitory effect on bacterial growth by inhibiting the expression of the following essential genes: inhA (a fatty acid elongase), rpsL (ribosomal S12 protein), gyrA (DNA gyrase), pncA (pyrazinamidase), polA (DNA polymerase I) and rpoC (RNA polymerase β subunit) of M. smegmatis. Each PNA was tested at 20 μM, 10 μM, 5 μM and 2.5 μM concentrations to determine whether they caused a dose dependent killing of M. smegmatis cultured in Middlebrook 7H9 broth or in a J774A.1 murine macrophage cell line. Results: In Middlebrook broth, the strong growth inhibitory effect against M. smegmatis was observed by PNAs targeting the inhA and rpsL genes at all four concentrations. The PNAs targeting the pncA, polA and rpoC genes were found to exhibit strong growth inhibition against M. smegmatis but only at 20 μM concentration. No growth inhibition of M. smegmatis was seen in pure culture when treated with PNAs targeting gyrA and a mismatch PNA targeting dnaG (DNA primase). All six PNAs showed killing of M. smegmatis in J774A.1 macrophage cell line that were statistically significant (p Conclusion: It may be concluded from this study that PNAs could be potential therapeutics for mycobacterial infections.

关键词： Middlebrook 7H9 Broth Culture J774A.1 Murine Macrophage Cell Line Antisense Therapy peptide nucleic acid Cell Penetrating peptide Mycobacterium

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Virus-like supramolecular assemblies formed by cooperation of base pairing interaction and peptidic association

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Science China Chemistry 2016年第3期59卷 310-315页

作者： Meiwen Cao Ningning Wang Peng Zhou Yawei Sun Jiqian Wang Shengjie Wang Hai Xu Centre for Bioengineering and Biotechnology State Key Laboratory of Heavy Oil ProcessingCollege of Chemical EngineeringChina University of Petroleum(East China)Qingdao266555China

A peptide nucleic acid （PNA）-peptide conjugated molecule, T＇3（AKAE）2, was designed to have both a PNA segment for oligo- nucleotide binding and an ionic self-complementary peptide sequence for self-association. T＇3（AKAE）2 could co-assemble with oligoadenines （d（A）x） to form virus-like supramolecular structures whose morphology showed dependence on the chain length and rigidity of the d（A）x molecules. Smaller nanospheres with diameters of 13.0±2.0 nm were produced in the case of d（A）6. Wormlike aggregates with lengths of 20-50 nm and diameters of 15.0±2.5 nm were found in the cases of d（A）12, d（A）ls, d（A）24 and d（A）30. And larger spherical aggregates with diameters of 18±5 nm came into presence in the cases of d（A）36 and d（A）42＋. These nanostructures were suggested to be formed under a cooperative effect of base pair recognition and peptidic association. The study provides insights into the programmed assembly of a multi-components system as well as control of the size and shade of the co-assembled structures, which is of great significance in develouing gene/drug deliverv systems.

关键词： peptide nucleic acid oligonucleotide artificial virus co-assembly supramolecular complex

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Synthesis of Peptoid nucleic acid with Thymine as nucleic Base

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Chinese Chemical Letters 2000年第9期11卷 771-774页

作者： Yun WU Jie Cheng XU Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032 Shanghai Inst. of Organic Chemistry Chinese Academy of Sciences Shanghai 200032 China

The synthesis of peptoid nucleic acid bearing thymine as nucleobase has been achieved. This modified oligonucleotide showed good hybridization with DNA.

关键词： peptoid peptide nucleic acid solid phase synthesis thymine

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