Short DNA Oligonucleotide as a Ag+ Binding Detector.
Ontology highlight
ABSTRACT:
SUBMITTER: Shen F
PROVIDER: S-EPMC7658945 | biostudies-literature | 2020 Nov
REPOSITORIES: biostudies-literature
Publications
Short DNA Oligonucleotide as a Ag<sup>+</sup> Binding Detector.
Shen Fusheng F Mao Song S Mathivanan Johnsi J Wu Ying Y Chandrasekaran Arun Richard AR Liu Hehua H Gan Jianhua J Sheng Jia J
ACS omega 20201028 44
Ag<sup>+</sup> has been known to mediate several natural metallo-base pairs. Based on the unique structural information of a short 8-mer DNA strand (5'-GCACGCGC-3') induced by Ag<sup>+</sup>, we constructed several fluorescent DNA beacons for the detection of Ag<sup>+</sup> according to the increase in the fluorescence emission on Ag<sup>+</sup> binding. This Ag<sup>+</sup> detection assay is quick, sensitive, and easy to adapt and can function in a wide range of temperatures from 5 to 65 °C. ...[more]
Similar Datasets
Project description:DNA/DNA duplex formation is the basic mechanism that is used in genome tiling arrays and SNP arrays manufactured by Affymetrix. However, detailed knowledge of the physical process is still lacking. In this study, we show a free energy analysis of DNA/DNA duplex formation these arrays based on the positional-dependent nearest-neighbor (PDNN) model, which was developed previously for describing DNA/RNA duplex formation on expression microarrays. Our results showed that the two ends of a probe contribute less to the stability of the duplexes and that there is a microarray surface effect on binding affinities. We also showed that free energy cost of a single mismatch depends on the bases adjacent to the mismatch site and obtained a comprehensive table of the cost of a single mismatch under all possible combination of adjacent bases. The mismatch costs were found to be correlated with those determined in aqueous solution. We further demonstrate that the DNA copy number estimated from the SNP array correlates negatively with the target length; this is presumably caused by inefficient PCR amplification for long fragments. These results provide important insights into the molecular mechanisms of microarray technology and have implications for microarray design and the interpretation of observed data.
| S-EPMC1807971 | biostudies-literature
Project description:Metallothioneins (MTs) constitute a family of cysteine-rich proteins that play key biological roles for a wide range of metal ions, but unlike many other metalloproteins, the structures of apo- and partially metalated MTs are not well understood. Here, we combine nano-electrospray ionization-mass spectrometry (ESI-MS) and nano-ESI-ion mobility (IM)-MS with collision-induced unfolding (CIU), chemical labeling using N-ethylmaleimide (NEM), and both bottom-up and top-down proteomics in an effort to better understand the metal binding sites of the partially metalated forms of human MT-2A, viz., Ag4-MT. The results for Ag4-MT are then compared to similar results obtained for Cd4-MT. The results show that Ag4-MT is a cooperative product, and data from top-down and bottom-up proteomics mass spectrometry analysis combined with NEM labeling revealed that all four Ag+ ions of Ag4-MT are bound to the β-domain. The binding sites are identified as Cys13, Cys15, Cys19, Cys21, Cys24, and Cys26. While both Ag+ and Cd2+ react with MT to yield cooperative products, i.e., Ag4-MT and Cd4-MT, these products are very different; Ag+ ions of Ag4-MT are located in the β-domain, whereas Cd2+ ions of Cd4-MT are located in the α-domain. Ag6-MT has been reported to be fully metalated in the β-domain, but our data suggest the two additional Ag+ ions are more weakly bound than are the other four. Higher order Agi-MT complexes (i = 7-17) are formed in solutions that contain excess Ag+ ions, and these are assumed to be bound to the α-domain or shared between the two domains. Interestingly, the excess Ag+ ions are displaced upon addition of NEM to this solution to yield predominantly Ag4NEM14-MT. Results from CIU suggest that Agi-MT complexes are structurally more ordered and that the energy required to unfold these complexes increases as the number of coordinated Ag+ increases.
| S-EPMC8114364 | biostudies-literature
Project description:MYC, originally named c-myc, is an oncogene deregulated in many different forms of cancer. Translocation of the MYC gene to an immunoglobulin gene leads to an overexpression and the development of Burkitt's lymphoma (BL). Sporadic BL constitutes one subgroup where one of the translocation sites is located at the 5'-vicinity of the two major MYC promoters P₁ and P₂. A non-B-DNA forming sequence within this region has been reported with the ability to form an intramolecular triplex (H-DNA) or a G-quadruplex. We have examined triplex formation at this site first by using a 17 bp triplex-forming oligonucleotide (TFO) and a double strand DNA (dsDNA) target corresponding to the MYC sequence. An antiparallel purine-motif triplex was detected using electrophoretic mobility shift assay. Furthermore, we probed for H-DNA formation using the BQQ-OP based triplex-specific cleavage assay, which indicated the formation of the structure in the supercoiled plasmid containing the corresponding region of the MYC promoter. Targeting non-B-DNA structures has therapeutic potential; therefore, we investigated their influence on strand-invasion of anti-gene oligonucleotides (ON)s. We show that in vitro, non-B-DNA formation at the vicinity of the ON target site facilitates dsDNA strand-invasion of the anti-gene ONs.
| S-EPMC6429085 | biostudies-other
Project description:G-quadruplexes (G4s) are well known non-canonical DNA secondary structures that can form in human cells. Most of the tools available to investigate G4-biology rely on small molecule ligands that stabilise these structures. However, the development of probes that disrupt G4s is equally important to study their biology. In this study, we investigated the disruption of G4s using Locked Nucleic Acids (LNA) as invader probes. We demonstrated that strategic positioning of LNA-modifications within short oligonucleotides (10 nts.) can significantly accelerate the rate of G4-disruption. Single-molecule experiments revealed that short LNA-probes can promote disruption of G4s with mechanical stability sufficient to stall polymerases. We corroborated this using a single-step extension assay, revealing that short LNA-probes can relieve replication dependent polymerase-stalling at G4 sites. We further demonstrated the potential of such LNA-based probes to study G4-biology in cells. By using a dual-luciferase assay, we found that short LNA probes can enhance the expression of c-KIT to levels similar to those observed when the c-KIT promoter is mutated to prevent the formation of the c-KIT1 G4. Collectively, our data suggest a potential use of rationally designed LNA-modified oligonucleotides as an accessible chemical-biology tool for disrupting individual G4s and interrogating their biological functions in cells.
| S-EPMC9303293 | biostudies-literature
Project description:Both in situ synthesized long oligo arrays from Agilent Technologies and short oligo arrays from Affymetrix were used to meausre differential gene expression in RNA samples generated from human neural stem cells treated with vehicle (EtOH, 0.01%) and tert-butylhydroquinone (tBHQ, 20µM) for 24h. For Affymetrix technology, RNAs from vehicle or tBHQ treated groups were analyzed separately. There are three replicates (GSM 11755, 11756, 11780, 11781, 11782, and 11783) which represent the RNA preps harvested at three different time (Jan 16, 2002, Jan 24, 2002, and Feb 20, 2002). For Agilent arrays, amplified cRNAs generated from vehicle or tBHQ treated groups were labeled with cy3 or cy5 and were competitively hybridized with Hu 1A oligo arrays. The same RNA preps used for Affymetrix array analysis were also used for Agilent array analysis. Finally, three replicates (GSM 11803, 11804 and 11809) were generated. Keywords: parallel sample
2003-10-22 | GSE759 | GEO
Project description:Here we describe the formation of an unexpected and unique family of hollow six-stranded helicates. The formation of these structures depends on the coordinative flexibility of silver and the 2-formyl-1,8-napthyridine subcomponent. Crystal structures show that these assemblies are held together by Ag4I, Ag4Br, or Ag6(SO4)2 clusters, where the templating anion plays an integral structure-defining role. Prior to the addition of the anionic template, no six-stranded helicate was observed to form, with the system instead consisting of a dynamic mixture of triple helicate and tetrahedron. Six-stranded helicate formation was highly sensitive to the structure of the ligand, with minor modifications inhibiting its formation. This work provides an unusual example of mutual stabilization between metal clusters and a self-assembled metal-organic cage. The selective preparation of this anisotropic host demonstrates new modes of guiding selective self-assembly using silver(I), whose many stable coordination geometries render design difficult.
| S-EPMC7879535 | biostudies-literature
Project description:BackgroundIn developing countries, death due to diseases caused by fecal-oral ingestion can be avoided by taking action on drinking water issues. Adequate access to water treatment systems to reduce infections is a critical cause. Silver has been used as an antibacterial product, including biomedical applications. Therefore, in this paper, the effect of the chemical speciation of silver from silver-modified zeolite-rich tuffs on the mortality of Escherichia coli (E. coli), Streptococcus faecalis (S. faecalis) and Candida albicans (C. albicans) suspended in aqueous solution was investigated for disinfection purposes.MethodsThe following aspects were considered to develop the investigation: a) the technique to prepare the modified zeolitic materials, either with ionic silver or silver nanoparticles, which were obtained in two ways: one, with grapefruit extract and the second, by using non-thermal plasma generated in a dielectric barrier discharge reactor of parallel plates; b) the response of the prokaryotes (bacteria) and eukaryote (yeast) microorganisms to disinfectant agents in batch systems; c) the disinfection processes as a function of time to obtain kinetics parameters; and d) the kinetics of the silver release from the silver-modified zeolite-rich tuffs, considering the models of Higuchi and Korsmeyer. The zeolitic materials were characterized by low-vacuum scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS).ResultsThe non-thermal plasma reduced ionic silver is more efficient at generating silver compounds with several oxidation states, which are essential during the microbial inhibition process. For the bacterial (E. coli and S. faecalis), the materials with nanoparticles were efficient to inactivate them. However, the yeast (C. albicans) reaches the total inactivation when the zeolitic material contains ionic silver in the crystalline network.ConclusionThe E. coli, S. faecalis and C. albicans survival behavior suspended in aqueous solutions after contact with Ag-modified natural zeolites depends on the chemical speciation of the silver present in these materials, Ag+1 in the case of OAgiZ or nanoparticles of Ago promoted by the grapefruit extract (OAgnpTZ), as well as by non-thermal plasma generated in a dielectric barrier discharge reactor of parallel plates (OAgnpPZ). In general, the concentration of silver in the aqueous solution after the disinfection process cannot exceed the recommended levels established for international organizations. The OAgnpPZ is a potential microbicide agent against E. coli and C. albicans, and the OAgn pTZ for F. faecalis.Graphical abstractARTWORK.
| S-EPMC7721831 | biostudies-literature
Project description:Both in situ synthesized long oligo arrays from Agilent Technologies and short oligo arrays from Affymetrix were used to measure differential gene expression in RNA samples generated from human neuroblastoma cells treated with vehicle (EtOH, 0.01%) and tert-butylhydroquinone (tBHQ, 10µM) for 24h. For Affymetrix technology, RNAs from vehicle or tBHQ treated groups were analyzed separately. There are three replicates (GSM 11865, 11866, 11858, 11857, 11870, and 11871) which represent the RNA preps harvested at three different time (April 17, 2001, March 15, 2001, and Feb 21, 2001). For Agilent arrays, amplified cRNAs generated from vehicle or tBHQ treated groups were labeled with cy3 or cy5 and were competitively hybridized with Hu 1A oligo arrays. The RNA prep used for Agilent array analysis was not the same as the one used for Affymetrix array analysis. Finally, three replicates (GSM 11828, 11831 and 11835) were generated. Keywords: parallel sample
2003-10-22 | GSE761 | GEO
Project description:Antisense oligonucleotide (ASO)-based therapy is one of the next-generation therapy, especially targeting neurological disorders. Many cases of ASO-dependent gene expression suppression have been reported. Recently, we developed a tocopherol conjugated DNA/RNA heteroduplex oligonucleotide (Toc-HDO) as a new type of drug. Toc-HDO is more potent, stable, and efficiently taken up by the target tissues compared to the parental ASO. However, the detailed mechanisms of Toc-HDO, including its binding proteins, are unknown. Here, we developed native gel shift assays with fluorescence-labeled nucleic acids samples extracted from mice livers. These assays revealed two Toc-HDO binding proteins, annexin A5 (ANXA5) and carbonic anhydrase 8 (CA8). Later, we identified two more proteins, apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) and flap structure-specific endonuclease 1 (FEN1) by data mining. shRNA knockdown studies demonstrated that all four proteins regulated Toc-HDO activity in Hepa1-6, mouse hepatocellular cells. In vitro binding assays and fluorescence polarization assays with purified recombinant proteins characterized the identified proteins and pull-down assays with cell lysates demonstrated the protein binding to the Toc-HDO and ASO in a biological environment. Taken together, our findings provide a brand new molecular biological insight as well as future directions for HDO-based disease therapy.
| S-EPMC8136785 | biostudies-literature
Project description:Owing to its great threat to human health and environment, Pb2+ pollution has been recognized as a major public problem by the World Health Organization (WHO). Many DNA aptamers have been utilized in the development of Pb2+-detection sensors, but the underlying mechanisms remain elusive. Here, we report three Pb2+-complexed structures of the thrombin binding aptamer (TBA). These high-resolution crystal structures showed that TBA forms intramolecular G-quadruplex and Pb2+ is bound by the two G-tetrads in the center. Compared to K+-stabilized G-quadruplexes, the coordinating distance between Pb2+ and the G-tetrads are much shorter. The T3T4 and T12T13 linkers play important roles in dimerization and crystallization of TBA, but they are changeable for Pb2+-binding. In combination with mutagenesis and CD spectra, the G8C mutant structure unraveled that the T7G8T9 linker of TBA is also variable. In addition to expansion of the Pb2+-binding aptamer sequences, our study also set up one great example for quick and rational development of other aptamers with similar or optimized binding activity.
| S-EPMC8748701 | biostudies-literature