共查询到20条相似文献,搜索用时 46 毫秒
1.
Dipl.‐Chem. Björn Niebel Christian Lentz Dipl.‐Biol. Monika Pofahl Prof. Dr. Günter Mayer Prof. Dr. Achim Hoerauf Dr. Kenneth M. Pfarr Prof. Dr. Michael Famulok 《Chemistry (Weinheim an der Bergstrasse, Germany)》2010,16(36):11100-11107
Functional nucleic acids, such as aptamers and allosteric ribozymes, can sense their ligands specifically, thereby undergoing structural alterations that can be converted into a detectable signal. The direct coupling of molecular recognition to signal generation enables the production of versatile reporters that can be applied as molecular probes for various purposes, including high‐throughput screening. Here we describe an unprecedented type of a nucleic acid‐based sensor system and show that it is amenable to high‐throughput screening (HTS) applications. The approach detects the displacement of an aptamer from its bound protein partner by means of luminescent oxygen channeling. In a proof‐of‐principle study we demonstrate that the format is feasible for efficient identification of small drug‐like molecules that bind to a protein target, in this case to the Sec7 domain of cytohesin. We extended the approach to a new cytohesin‐specific single chain DNA aptamer, C10.41, which exhibits a similar binding behavior to cytohesins but has the advantage of being more stable and easier to synthesize and to modify than the RNA‐aptamer M69. The results obtained with both aptamers indicate the general suitability of the aptamer‐displacement assay based on luminescent oxygen channelling (ADLOC) for HTS. We also analyzed the potential for false positive hits and identified from a library of 18 000 drug‐like small molecules two compounds as strong singlet‐oxygen quenchers. With full automation and the use of commercially available plate readers, we estimate that the ADLOC‐based assay described here could be used to screen at least 100 000 compounds per day. 相似文献
2.
SDS‐PAGE procedure: Application for characterization of new entirely uncharged nucleic acids analogs
《Electrophoresis》2018,39(4):670-674
SDS‐PAGE is considered to be a universal method for size‐based separation and analysis of proteins. In this study, we applied the principle of SDS‐PAGE to the analysis of new entirely uncharged nucleic acid (NA) analogues, – phosphoryl guanidine oligonucleotides (PGOs). The procedure was also shown to be suitable for morpholino oligonucleotides (PMOs) and peptide nucleic acids (PNAs). It was demonstrated that SDS can establish hydrophobic interactions with these types of synthetic NAs, giving them a net negative charge and thus making these molecules mobile in polyacrylamide slab gels under the influence of an electric field. 相似文献
3.
Alexander L. Prinzen Daniel Saliba Christopher Hennecker Tuan Trinh Anthony Mittermaier Hanadi F. Sleiman 《Angewandte Chemie (International ed. in English)》2020,59(31):12900-12908
Triggering the release of small molecules in response to unique biomarkers is important for applications in drug delivery and biodetection. Due to low quantities of biomarker, amplifying release is necessary to gain appreciable responses. Nucleic acids have been used for both their biomarker‐recognition properties and as stimuli, notably in amplified small‐molecule release by nucleic‐acid‐templated catalysis (NATC). The multiple components and reversibility of NATC, however, make it difficult to apply in vivo. Herein, we report the use of the hybridization chain reaction (HCR) for the amplified, conditional release of small molecules from standalone nanodevices. We couple HCR with a DNA‐templated reaction resulting in the amplified, immolative release of small molecules. We integrate the HCR components into single nanodevices as DNA tracks and spherical nucleic acids, spatially isolating reactive groups until triggering. This could be applied to biosensing, imaging, and drug delivery. 相似文献
4.
Within the last decades we witnessed the discovery of a number of mechanisms that enable the use of nucleic acids for therapeutic purposes. Small RNA and DNA molecules can be used to specifically suppress the expression of individual genes. Aptamers provide an alternative to monoclonal antibodies. A prerequisite for the pharmacological use of nucleic acids is an enhanced stability towards the body's degrading enzymes. This can be achieved for instance by employing non‐natural mirror‐image nucleic acids. The article describes the basic principles of stereochemistry underlying this approach and shows how these translate into the discovery of mirror‐image aptamers. Furthermore, it explains why the stereospecificity of Watson‐Crick base pairing has precluded mirror‐image nucleic acids from gene silencing methods and introduces a new approach that may help to overcome this. 相似文献
5.
Michael Famulok Jack W. Szostak 《Angewandte Chemie (International ed. in English)》1992,31(8):979-988
In vitro selection is a method that allows the simultaneous screening of very large numbers of nucleic acid molecules for a wide range of properties from binding characteristics to catalytic properties; moreover, the isolation of the very rare functional molecules becomes possible. Binding sites between proteins and nucleic acids, for example, have been evaluated by this methodology in order to gain information about protein/nucleic acid interactions. Structure and function of catalytic RNA (“ribozymes”) has been studied by in vitro selection and has led to new ribozymes with improved catalytic function. Substrate specificity of catalytic RNA has been changed and has led to a ribozyme that cleaves DNA. Other applications include the isolation of nucleic acids that bind specifically to small organic molecules and of RNA molecules that form triple helices with double-stranded DNA. In this article we discuss the background, design, and results of in vitro genetic experiments, which bridge biochemical/molecular biological and organic chemical approaches to molecular recognition. 相似文献
6.
Qiu-Long Zhang Yang Wang Liang-Liang Wang Fan Xie Ruo-Yue Wu Xu-Yang Ma Han Li Yan Liu Prof. Shunchun Yao Prof. Liang Xu 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2023,135(2):e202214698
Nucleic acid (NA) computation has been widely developed in the past years to solve kinds of logic and mathematic issues in both information technologies and biomedical analysis. However, the difficulty to integrate non-NA molecules limits its power as a universal platform for molecular computation. Here, we report a versatile prototype of hybridized computation integrated with both nucleic acids and non-NA molecules. Employing the conformationally controlled ligand converters, we demonstrate that non-NA molecules, including both small molecules and proteins, can be computed as nucleic acid strands to construct the circuitry with increased complexity and scalability, and can be even programmed to solve arithmetical calculations within the computational nucleic acid system. This study opens a new door for molecular computation in which all-NA circuits can be expanded with integration of various ligands, and meanwhile, ligands can be precisely programmed by the nuclei acid computation. 相似文献
7.
One of the fundamental challenges in studying biomacromolecules (e.g. nucleic acids and proteins) and their complexes in a biological system is isolating them in their structurally and functionally intact forms. Electrophoresis offers convenient and efficient separation and analysis of biomacromolecules but recovery of separated biomacromolecules is a significant challenge. In this study, DNAs of various sizes were separated by electrophoresis in an acid‐degradable polyacrylamide gel. Almost 100% of the nucleic acids were recovered after the identified gel bands were hydrolyzed under a mildly acidic condition and purified using anion exchange resin. Further concentration by centrifugal filtration and a second purification using ion exchange column chromatography yielded 44–84% of DNA. The second conventional (non‐degradable) gel electrophoresis confirmed that the nucleic acids recovered from acid‐degradable gel bands preserved their electrophoretic properties through acidic gel hydrolysis, purification, and concentration processes. The plasmid DNA recovered from acid‐degradable gel transfected cells significantly more efficiently than the starting plasmid DNA (i.e. improved biological activity via acid‐degradable PAGE). Separation of other types of nucleic acids such as small interfering RNA using this convenient and efficient technique was also demonstrated. 相似文献
8.
Mattia Riccardo Monaco Belén Poladura Dr. Miriam Diaz de Los Bernardos Markus Leutzsch Dr. Richard Goddard Prof. Dr. Benjamin List 《Angewandte Chemie (International ed. in English)》2014,53(27):7063-7067
Organocatalysis, catalysis using small organic molecules, has recently evolved into a general approach for asymmetric synthesis, complementing both metal catalysis and biocatalysis. 1 Its success relies to a large extent upon the introduction of novel and generic activation modes. 2 Remarkably though, while carboxylic acids have been used as catalyst directing groups in supramolecular transition‐metal catalysis, 3 a general and well‐defined activation mode for this useful and abundant substance class is still lacking. Herein we propose the heterodimeric association of carboxylic acids with chiral phosphoric acid catalysts as a new activation principle for organocatalysis. This self‐assembly increases both the acidity of the phosphoric acid catalyst and the reactivity of the carboxylic acid. To illustrate this principle, we apply our concept in a general and highly enantioselective catalytic aziridine‐opening reaction with carboxylic acids as nucleophiles. 相似文献
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10.
Marcos V. Foguel Angelica M. Balcarcel Mark A. Reed Percy Calvo‐Marzal Yulia V. Gerasimova Dmitry M. Kolpashchikov Karin Y. Chumbimuni‐Torres 《Electroanalysis》2020,32(4):835-841
One major challenge in nucleic acids analysis by hybridization probes is a compromise between the probe's tight binding and sequence‐selective recognition of nucleic acid targets folded into stable secondary structures. We have been developing a four‐way junction (4WJ)‐based sensor that consists of a universal stem‐loop (USL) probe immobilized on an electrode surface and two adaptor strands (M and F). The sensor was shown to be highly selective towards single base mismatches at room temperature, able to detect multiple targets using the same USL probe, and have improved ability to detect folded nucleic acids. However, some nucleic acid targets, including natural RNA, are folded into very stable secondary and tertiary structures, which may represent a challenge even for the 4WJ sensors. This work describes a new sensor, named MVF since it uses three probe stands M, V and F, which further improves the performance of 4WJ sensors with folded targets. The MVF sensor interrogating a 16S rRNA NASBA amplicon with calculated folding energy of ?32.82 kcal/mol has demonstrated 2.5‐fold improvement in a signal‐to‐background ratio in comparison with a 4WJ sensor lacking strand V. The proposed design can be used as a general strategy in the analysis of folded nucleic acids including natural RNA. 相似文献
11.
Anke Dierckx Dr. Francois‐Alexandre Miannay Dr. Nouha Ben Gaied Søren Preus Markus Björck Prof. Dr. Tom Brown Prof. Dr. L. Marcus Wilhelmsson 《Chemistry (Weinheim an der Bergstrasse, Germany)》2012,18(19):5987-5997
Fluorescent‐base analogues (FBAs) comprise a group of increasingly important molecules for the investigation of nucleic acid structure and dynamics as well as of interactions between nucleic acids and other molecules. Here, we report on the synthesis, detailed spectroscopic characterisation and base‐pairing properties of a new environment‐sensitive fluorescent adenine analogue, quadracyclic adenine (qA). After developing an efficient route of synthesis for the phosphoramidite of qA it was incorporated into DNA in high yield by using standard solid‐phase synthesis procedures. In DNA qA serves as an adenine analogue that preserves the B‐form and, in contrast to most currently available FBAs, maintains or even increases the stability of the duplex. We demonstrate that, unlike fluorescent adenine analogues, such as the most commonly used one, 2‐aminopurine, and the recently developed triazole adenine, qA shows highly specific base‐pairing with thymine. Moreover, qA has an absorption band outside the absorption of the natural nucleobases (>300 nm) and can thus be selectively excited. Upon excitation the qA monomer displays a fluorescence quantum yield of 6.8 % with an emission maximum at 456 nm. More importantly, upon incorporation into DNA the fluorescence of qA is significantly less quenched than most FBAs. This results in quantum yields that in some sequences reach values that are up to fourfold higher than maximum values reported for 2‐aminopurine. To facilitate future utilisation of qA in biochemical and biophysical studies we investigated its fluorescence properties in greater detail and resolved its absorption band outside the DNA absorption region into distinct transition dipole moments. In conclusion, the unique combination of properties of qA make it a promising alternative to current fluorescent adenine analogues for future detailed studies of nucleic acid‐containing systems. 相似文献
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Dr. Céline Douat Dr. Christopher Aisenbrey Stéphanie Antunes Dr. Marion Decossas Dr. Olivier Lambert Dr. Burkhard Bechinger Dr. Antoine Kichler Dr. Gilles Guichard 《Angewandte Chemie (International ed. in English)》2015,54(38):11133-11137
Despite significant advances in foldamer chemistry, tailored delivery systems based on foldamer architectures, which provide a high level of control over secondary structure, are curiously rare among non‐viral technologies for transporting nucleic acids into cells. A potent pH‐responsive, bioreducible cell‐penetrating foldamer (CPF) was developed through covalent dimerization of a short (8‐mer) amphipathic oligourea sequence bearing histidine‐type units. This CPF exhibits a high capacity to assemble with pDNA and mediates efficient delivery of nucleic acids into the cell. Furthermore, it does not adversely affect cellular viability and was shown to compare favorably with a cognate peptide transfection agent based on His‐rich sequences. 相似文献
14.
Kamyar Yazdani Deondre Jordan Mo Yang Christopher R. Fullenkamp David R. Calabrese Robert Boer Thomas Hilimire Timothy E. H. Allen Rabia T. Khan John S. Schneekloth Jr. 《Angewandte Chemie (International ed. in English)》2023,62(11):e202211358
Small molecule targeting of RNA has emerged as a new frontier in medicinal chemistry, but compared to the protein targeting literature our understanding of chemical matter that binds to RNA is limited. In this study, we reported R epository O f BI nders to N ucleic acids (ROBIN), a new library of nucleic acid binders identified by small molecule microarray (SMM) screening. The complete results of 36 individual nucleic acid SMM screens against a library of 24 572 small molecules were reported (including a total of 1 627 072 interactions assayed). A set of 2 003 RNA-binding small molecules was identified, representing the largest fully public, experimentally derived library of its kind to date. Machine learning was used to develop highly predictive and interpretable models to characterize RNA-binding molecules. This work demonstrates that machine learning algorithms applied to experimentally derived sets of RNA binders are a powerful method to inform RNA-targeted chemical space. 相似文献
15.
Bridging the Two Worlds: A Universal Interface between Enzymatic and DNA Computing Systems 下载免费PDF全文
Shay Mailloux Dr. Yulia V. Gerasimova Dr. Nataliia Guz Dr. Dmitry M. Kolpashchikov Prof. Evgeny Katz 《Angewandte Chemie (International ed. in English)》2015,54(22):6562-6566
Molecular computing based on enzymes or nucleic acids has attracted a great deal of attention due to the perspectives of controlling living systems in the way we control electronic computers. Enzyme‐based computational systems can respond to a great variety of small molecule inputs. They have the advantage of signal amplification and highly specific recognition. DNA computing systems are most often controlled by oligonucleotide inputs/outputs and are capable of sophisticated computing as well as controlling gene expressions. Here, we developed an interface that enables communication of otherwise incompatible nucleic‐acid and enzyme‐computational systems. The enzymatic system processes small molecules as inputs and produces NADH as an output. The NADH output triggers electrochemical release of an oligonucleotide, which is accepted by a DNA computational system as an input. This interface is universal because the enzymatic and DNA computing systems are independent of each other in composition and complexity. 相似文献
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Phosphine‐Free Stille–Migita Chemistry for the Mild and Orthogonal Modification of DNA and RNA 下载免费PDF全文
Dipl.‐Biochem. André Krause Alexander Hertl Fabian Muttach Prof. Dr. Andres Jäschke 《Chemistry (Weinheim an der Bergstrasse, Germany)》2014,20(50):16613-16619
An optimized catalyst system of [Pd2(dba)3] and AsPh3 efficiently catalyzes the Stille reaction between a diverse set of functionalized stannanes and halogenated mono‐, di‐ and oligonucleotides. The methodology allows for the facile conjugation of short and long nucleic acid molecules with moieties that are not compatible with conventional chemical or enzymatic synthesis, among them acid‐, base‐, or fluoride‐labile protecting groups, fluorogenic and synthetically challenging moieties with good to near‐quantitative yields. Notably, even azides can be directly introduced into oligonucleotides and (deoxy)nucleoside triphosphates, thereby giving direct access to “clickable” nucleic acids. 相似文献
18.
Herein, we reported a cationic conjugated polymers-based new biosensor with label-free and fluorescence turn-on strategy by virtue of targets regulated aggregation and quenching ability of perylene diimide derivatives. 相似文献
19.
Two‐dimensional (2D) layered nanomaterials, e.g. graphene and molybdenum disulfide (MoS2), have rapidly emerged in material sciences due to their unique physical, chemical and mechanical properties. In the meanwhile, there is a growing interest in constructing electrochemical sensors for a wide range of chemical and biological molecules by using these 2D nanomaterials. In this review, we summarize recent advances on using graphene and MoS2 for the development of electrochemical sensors for small molecules, proteins, nucleic acids and cells detection. We also provide our perspectives in this rapidly developing field. 相似文献
20.
Here, we report the evolution of two star-shaped (five-way junction) deoxyribozymes from a catalytic DNA containing a three-way junction scaffold. The transition was shown to be a switch rather than a gradual progression. The star-shaped motifs, surprisingly, only took five selection cycles to be detected, and another four to dominate the evolving population. Chemical probing experiments indicated that the two deoxyribozymes belong to the same family despite noticeable variations in both the primary sequence and the secondary structure. Our findings not only describe the evolution of high-branching nucleic acid structures from a low-branching catalytic module, but they also illustrate the idea of deriving a rare structural motif by sampling the sequence variants of a given functional nucleic acid. 相似文献