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CHEMICAL RESEARCH IN CHINESE UNIVERSITIES · 2020 ·Vol. 36 ·期 2 ·301-306 · IF 1.063

Programmable and Reversible Regulation of Catalytic Hemin@MOFs Activities with DNA Structures

Liu Shuo,Yang Mingjie,Guo Weiwei

Metal-organic frameworks(MOFs)-based nanozyme plays an important role in biosensing, therapy and catalysis. In this study, the effects of single-stranded DNA(ssDNA) with programmable sequences and its complementary DNA(TDNA) on the intrinsic peroxidase-like activity of hemin loaded MOFs(UiO-66- 2 ), denoted as hemin@UiO-66-NH 2 , were investigated. The hemin@UiO-66- 2 exhibited improved catalytic activity compared with free hemin. However, the catalytic activity is inhibited in the presence of ssDNA, as ssDNA can be adsorbed by MOFs and therefore protected the active sites from contact with substrates. Upon the addition of the T DNA , double-stranded DNA(dsDNA) was formed and detached from the MOFs, resulting in the recovery of catalytic activity. Sequentially adding ssDNA or its complementary DNA strands can achieve the reversible regulation of the catalytic activity of MOFs nanozymes. Moreover, the DNA hybridization-based regulation was further applied to a cascaded catalytic system composed of the nanozyme, hemin@UiO-66-NH 2 , and glucose oxidase. These nanozyme based programmable and reversibly regulated catalytic systems may have potential applications in future smart biosensing and catalysis systems.

研究领域: 生物化学 · 纳米医学 · 催化科学
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CELL · 2019 · IF 36.216

Acetylation Blocks cGAS Activity and Inhibits Self-DNA-Induced Autoimmunity

Jiang Dai,Yi-Jiao Huang,Xinhua He,Ming Zhao,Xinzheng Wang,Zhao-Shan Liu,Wen Xue,Hong Cai,Xiao-Yan Zhan,Shao-Yi Huang,Kun He,Hongxia Wang,Na Wang,Zhihong Sang,Tingting Li,Qiu-Ying Han,Jie Mao,Xinwei Diao,Nan Song,Yuan Chen,Wei-Hua Li,Jiang-Hong Man,Ai-Ling Li,Tao Zhou,Zheng-Gang Liu,Xue-Min Zhang,Tao Li

Summary The presence of DNA in the cytoplasm is normally a sign of microbial infections and is quickly detected by cyclic GMP-AMP synthase (cGAS) to elicit anti-infection immune responses. However, chronic activation of cGAS by self-DNA leads to severe autoimmune diseases for which no effective treatment is available yet. Here we report that acetylation inhibits cGAS activation and that the enforced acetylation of cGAS by aspirin robustly suppresses self-DNA-induced autoimmunity. We find that cGAS acetylation on either Lys384, Lys394, or Lys414 contributes to keeping cGAS inactive. cGAS is deacetylated in response to DNA challenges. Importantly, we show that aspirin can directly acetylate cGAS and efficiently inhibit cGAS-mediated immune responses. Finally, we demonstrate that aspirin can effectively suppress self-DNA-induced autoimmunity in Aicardi-Goutières syndrome (AGS) patient cells and in an AGS mouse model. Thus, our study reveals that acetylation contributes to cGAS activity regulation and provides a potential therapy for treating DNA-mediated autoimmune diseases.

研究领域: 免疫学 · 分子生物学 · 药理学
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ACS Applied Materials & Interfaces · 2016 · IF 7.145

Self-Assembled DNA Hydrogel Based on Enzymatically Polymerized DNA for Protein Encapsulation and Enzyme/DNAzyme Hybrid Cascade Reaction

Binbin Xiang,Kaiyu He,Rong Zhu,Zhuoliang Liu,Shu Zeng,Yan Huang,Zhou Nie,Shouzhuo Yao

DNA hydrogel is a promising biomaterial for biological and medical applications due to its native biocompatibility and biodegradability. Herein, we provide a novel, versatile, and cost-effective approach for self-assembly of DNA hydrogel using the enzymatically polymerized DNA building blocks. The X-shaped DNA motif was elongated by terminal deoxynucleotidyl transferase (TdT) to form the building blocks, and hybridization between dual building blocks via their complementary TdT-polymerized DNA tails led to gel formation. TdT polymerization dramatically reduced the required amount of original DNA motifs, and the hybridization-mediated cross-linking of building blocks endows the gel with high mechanical strength. The DNA hydrogel can be applied for encapsulation and controllable release of protein cargos (for instance, green fluorescent protein) due to its enzymatic responsive properties. Moreover, this versatile strategy was extended to construct a functional DNAzyme hydrogel by integrating the peroxidase-mimicking DNAzyme into DNA motifs. Furthermore, a hybrid cascade enzymatic reaction system was constructed by coencapsulating glucose oxidase and β-galactosidase into DNAzyme hydrogel. This efficient cascade reaction provides not only a potential method for glucose/lactose detection by naked eye but also a promising modular platform for constructing a multiple enzyme or enzyme/DNAzyme hybrid system.

研究领域: 生物材料学 · 分子生物学
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JACS Au · 2024 · IF 8.5

Leveraging Concentration Imbalance-Driven DNA Circuit as an Operational Amplifier to Enhance the Sensitivity of Hepatitis B Virus DNA Detection with Hybridization-Responsive DNA-Templated Silver Nanoclusters

Suo Lv,Qunyan Yao,Jiasheng Yi,Jingyi Si,Yifan Gao,Shao Su,Changfeng Zhu

Hepatitis B virus (HBV) infection remains a major global health concern, necessitating the development of sensitive and reliable diagnostic methods. In this study, we propose a novel approach to enhance the sensitivity of HBV DNA detection by leveraging a concentration imbalance-driven DNA circuit (CIDDC) as an operational amplifier, coupled with a hybridization-responsive DNA-templated silver nanocluster (DNA-AgNCs) nanoprobe named Q·C6-AgNCs. The CIDDC system effectively converts and amplifies the input HBV DNA into an enriched generic single-stranded DNA output, which subsequently triggers the fluorescence of the DNA-AgNCs reporter upon hybridization, generating a measurable signal for detection. By incorporating the DNA circuit, we not only achieved enhanced sensitivity with a lower detection limit of 0.11 nM but also demonstrated high specificity with single-base mismatch discriminability for HBV DNA detection. Additionally, this mix-and-detect assay format is simple, user-friendly, and isothermal. This innovative strategy holds promise for advancing molecular diagnostics and facilitating the effective management of HBV-related diseases.

CAS / MDL: 16940-66-2
研究领域: 分子生物学 · 医学诊断 · 纳米技术
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LUMINESCENCE · 2012 ·Vol. 28 ·期 6 ·860-864 · IF 1.731

Steady-state and time-resolved fluorescence of tetramethylrhodamine attached to DNA: correlation with DNA sequences

Tingting Chen,Leixiaomeng Fu,Lily Zu

Time-resolved fluorescence as well as steady-state absorption and fluorescence were detected in order to study the interactions between tetramethylrhodamine (TAMRA) and DNA when TAMRA was covalently labeled on single- and double-stranded oligonucleotides. Fluorescence intensity quenching and lifetime changes were characterized and correlated with different DNA sequences. The results demonstrated that the photoinduced electron transfer interaction between guanosine residues and TAMRA introduced a short lifetime fluorescence component when guanosine residues were at the TAMRA-attached terminal of the DNA sequences. The discrepancy of two-state and three-state models in previous studies was due to the DNA sequence selection and sensitivity of techniques used to detect the short lifetime component. The results will help the design of fluorescence-based experiments related to a dye labeled probe. Copyright © 2012 John Wiley & Sons, Ltd.

CAS / MDL: 77-86-1
研究领域: 化学 · 分子生物学 · 生物物理
✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS · 2019 · IF 2.983

Simultaneous quantitation of 14 DNA alkylation adducts in human liver and kidney cells by UHPLC-MS/MS: Application to profiling DNA adducts of genotoxic reagents

Yu Tang,Zhe Wang,Menglin Li,Ruiping Zhang,Jinlan Zhang

A rapid, sensitive and wide coverage ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method has been developed and validated for the simultaneous quantitation of 14 alkylation DNA adducts in cell genomic DNA, RNA and cell contents isolated from the in vitro cultured human kidney cell line 293 T and the human liver cell line L02 exposed to 3 genotoxic reagents: N -methyl -N- nitrosourea (MNU), methyl methanesulfonate (MMS) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). After exposure, DNA was isolated and directly hydrolysed under acid conditions or digested by enzymes to obtain the hydrolysates containing DNA alkylation adducts followed by optimization of the pretreatment method and chromatographic separation conditions. Quantification was performed on a Waters ACQUITY UPLC BEH Amide column (1.7 μm, 2.1 × 150 mm) using an electrospray ionization (ESI) source in positive mode by selective reaction monitoring (SRM) at the precursor to product ion transitions of 14 analytes. The method showed selectivity, good linearity (r>0.9950), accuracy (82.1%–115%), and intra-day (RSD%<14%) and inter-day (RSD%<15%) precision for 14 analytes. The recoveries of two pretreatment methods were all more than 50.5%, and no relative matrix effects were observed. Additionally, the samples were stable after short-term storage at 20 ℃ for 2 h, at 4 ℃ for 48 h or one cycle of freeze-thaw at −80 ℃. The established UHPLC-MS/MS method was used to evaluate the changes in alkylation DNA adducts and epigenetic modification-related methylcytosine after exposure to genotoxic reagents. For the first time, the results demonstrated that 3 genotoxic reagents induced different total amounts of adducts in the following sequence: MMS > NNK > MNU, and showed significant differences in the ratios of 7MeG to 1MeA and 1MeG to 1MeA in the 293 T cell model. Meanwhile, 293 T and L02 cells revealed significantly different DNA adduct formation characteristics in the contents of 1MeG and 1MeA. The DNA adduct formation relationships between DNA, RNA, and cell contents were probed to predict cancer risk and potential genotoxic exposure. This approach could be used to investigate the DNA adducts, their formation and the relationship to the mutagenicity or carcinogenicity of genotoxic reagents in future studies.

研究领域: 毒理学 · 分析化学 · 分子生物学
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MICROCHEMICAL JOURNAL · 2021 · IF 4.821

Metal-bridged DNA-functionalized silica nanoparticles for multifacet biological applications

Yue Huang,Jiehua Ma,Yuting Yan,Chao Li

Silicon, as the second richest element in the earth, is of great importance to the current nanoscience. Functionalization of silica nanomaterials with DNA molecules can significantly broaden their biological application. However, current assembly methods usually involve complex and time-consuming steps or detrimental reagents, limiting their real application. In this work, we report a metal ion-assistant DNA assembly method to simply, rapidly and robustly immobilize short DNA molecules on the surface of silica nanoparticles (SiNPs) and use it to perform different tasks. By fully screening the metal ions, metal ions such as Zn 2+ , Al 3+ , Y 3+ and Zr 4+ , are found to be effective on the mediation of the interaction between DNA and SiNPs. The DNA density can be finely tuned by adjusting the added concentration of DNA or metal ions, and the adsorbed DNA is highly stable in the metal ion-containing buffer. Through simply adjusting the kind of metal ions and DNA, different biological applications such as biosensing, purification, and gene regulation can be achieved. This work not only offers fundamental insights into the biointerface science of SiNPs with DNA, but holds great potential in the area of biomolecule purification, nucleic acid delivery, biosensing, and bioimaging.

研究领域: 纳米技术 · 生物技术 · 生物化学 · 材料科学
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Journal of Materials Chemistry B · 2022 ·Vol. 10 ·期 9 ·1386-1392 · IF 7.571

High-sensitivity fluorescence detection for lung cancer CYFRA21-1 DNA based on accumulative hybridization of quantum dots

Ling Yang,Ping’an Ma,Xiaorui Chen,Ziyong Cheng,Jun Lin

Sensitive detection of circulating tumor DNA (ctDNA) in vitro has attracted growing attention owing to its potential application in diagnostics of cancer. In this study, we synthesized hydrophilic AgInS2@ZnS core–shell quantum dot nanocrystals and magnetic Fe3O4 nanoparticles, and then the ctDNA triggered hybridization chain reaction was used to detect the CYFRA21-1 DNA associated with lung cancer. In the presence of CYFRA21-1 DNA, three hairpin structures were activated to turn on successively, resulting in the accumulation of quantum dots and eliciting considerable changes of the fluorescence signal. Compared with the conventional fluorescence detection, Fe3O4 provides magnetic adsorption properties and a large surface area for immobilizing and aggregating quantum dot nanoparticles attached to single-stranded DNA. The concentration of CYFRA21-1 is closely related to the number of quantum dots remaining after magnetic adsorption, which provides a promising approach for ctDNA quantification.

研究领域: 生物化学 · 分子诊断 · 纳米医学
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MICROCHEMICAL JOURNAL · 2023 · IF 4.8

CRISPR/Cas12a coupled with enzyme-DNA molecular switch photoelectrochemical assay for HIV nucleic acid

Lilin Qin,Fangxu Lou,Yan Wang,Yinhao Zhang,Shishi Liu,Xu Hun

Molecular switches formed by enzyme-DNA hybrid compounds combined with CRISPR-Cas12a cis -cleavage used for photoelectrochemical nucleic acid assay was fabricated. A compound, molecular switch, consisting of Taq DNA polymerase, aptamer, block DNA and methylene blue labeled primer (primer-Mb) is formed. Block DNA is assembled on the surface of MBs (block DNA-MBs), and hybridized with the primer-Mb. In the presence of Human Immunodeficiency Virus (HIV) target, molecular switches are turned on by hybridization of the target with block DNA. The Taq DNA polymerase is released and activated. The activated Taq DNA polymerase elongates the primer via the template, block DNA, which attached onto Magnetic beads (MBs). A dsDNA with Mb (dsDNA-Mb) was formed. With the assistance of Cas12a, a amplified photoelectrochemical signal was obtained and the HIV target DNA was qualified. Under the best conditions, the limit of detection (LOD) can be as low as 0.3 fM. This photoelectrochemical assay extends the tool kit of molecular nanotechnology.

CAS / MDL: 25952-53-8 6066-82-6
研究领域: 分子生物学 · 生物化学 · 纳米技术 · 病毒学
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ACS Sustainable Chemistry & Engineering · 2024 · IF 7.1

Preparation of Biomass-Derived DNA-Chitosan Polyelectrolyte Complex Hydrogels for Economical and Reliable DNA Preservation and Protein Immobilization

Yuhui Du,Xinqian Zheng,Yuan Ma,Fangjun Ge,Qiuyan Li,Hao Xiang,Xueyi Chen,Hai Zhou,Taolin Sun,Tingjian Chen

DNA-based biomaterials, including polyelectrolyte complex (PEC) hydrogels composed of DNA and other molecules, have received considerable attention due to their programmability and good biocompatibility. However, the cost for DNA synthesis is fairly high, especially for applications where a large quantity of DNA is needed. Construction of biomaterials using biomass-derived DNA is a feasible strategy to address this challenge. In this work, we fabricated and characterized PEC hydrogels with or without covalent cross-linking using biomass-derived DNA and chitosan (CS). Characterization results revealed detailed microstructures and mechanical properties of these two hydrogels, suggesting their potential application in DNA preservation and protein immobilization. We next developed a versatile DNA preservation strategy combining DNA-CS PEC hydrogel-based DNA encapsulation and chitosanase-mediated DNA release, which successfully maintained the integrity and functionality of various types of DNA, i.e., double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), plasmid DNA, and genomic DNA, during the treatment of DNase I and hydrogen peroxide (H2O2). Remarkably, this method also demonstrated good efficiency in protecting DNA containing a broadly used unnatural base pair, dNaM-dTPT3, against the attack of silver ion. Additionally, we developed a convenient and reliable strategy for protein immobilization with the covalently cross-linked PEC hydrogel. This strategy demonstrated good performance in the reusability of the immobilized enzyme, evidenced by 56.4% product yield of the initial cycle of usage achieved by the immobilized horseradish peroxidase (HRP) in the tenth cycle of usage. This work not only broadens the application scope of biomass-derived polymers, but also provides feasible strategies for the sustainable and cost-effective preservation and immobilization of biological macromolecules.

研究领域: 生物材料 · 生物技术 · 纳米技术 · 生物化学 · 分子生物学
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ACS Sensors · 2024 · IF 8.2

Label-Free Detection of DNA Base Mutation and Hybridized DNA by an Electrostatically Modified 3D Plasmonic Array

Xiaolu Xing,Yuhao Zhao,Haiyan Zhao,Ying Zhang,Ziqiu Chen,Bowen Liu,Bin Ren

Surface-enhanced Raman scattering (SERS) represents a promising avenue for DNA detection as it offers intrinsic chemical insights with high sensitivity compared to conventional methods. However, label-free and quantitative detection of unmodified DNA by SERS remains a major challenge in DNA analysis. To overcome this challenge, we propose a positively charged plasmonic nanosurface for DNA capture and quantitative analysis. Highly sensitive and uniform SERS enhancement was realized by a three-dimensional plasmonic array supporting well-designed hybrid plasmonic modes. Subsequently, the plasmonic array was modified with an electrostatically functionalized PDDA (poly(diene-dimethylammonium-chloride)) self-assembled monolayer in a single step. The effectiveness of the resulting PDDA-SERS substrate was demonstrated by the label-free and quantitative detection of base content and base mutation in hybridized DNA. The PDDA-SERS substrate provides a robust platform for SERS analysis not only of DNA but also of other electronegative analytes.

CAS / MDL: 569-64-2
研究领域: 生物化学 · 纳米技术 · 分析化学
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NUCLEIC ACIDS RESEARCH · 2019 ·Vol. 47 ·期 22 ·11514-11526 · IF 11.147

Sulfur-centered hemi-bond radicals as active intermediates in S-DNA phosphorothioate oxidation

Jie Jialong,Xia Ye,Huang Chun-Hua,Zhao Hongmei,Yang Chunfan,Liu Kunhui,Song Di,Zhu Ben-Zhan,Su Hongmei

Phosphorothioate (PS) modifications naturally appear in bacteria and archaea genome and are widely used as antisense strategy in gene therapy. But the chemical effects of PS introduction as a redox active site into DNA (S-DNA) is still poorly understood. Herein, we perform time-resolved spectroscopy to examine the underlying mechanisms and dynamics of the PS oxidation by potent radicals in free model, in dinucleotide, and in S-oligomer. The crucial sulphur-centered hemi-bonded intermediates -P–S∴S–P- were observed and found to play critical roles leading to the stable adducts of -P–S–S–P-, which are backbone DNA lesion products. Moreover, the oxidation of the PS moiety in dinucleotides d[GPSG], d[APSA], d[GPSA], d[APSG] and in S-oligomers was monitored in real-time, showing that PS oxidation can compete with adenine but not with guanine. Significantly, hole transfer process from A+• to PS and concomitant -P–S∴S–P- formation was observed, demonstrating the base-to-backbone hole transfer unique to S-DNA, which is different from the normally adopted backbone-to-base hole transfer in native DNA. These findings reveal the distinct backbone lesion pathway brought by the PS modification and also imply an alternative -P–S∴S–P-/-P–S–S–P- pathway accounting for the interesting protective role of PS as an oxidation sacrifice in bacterial genome.

研究领域: 生物化学 · 分子生物学 · 基因治疗
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