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Clickable PNA Probes for Imaging Human Telomeres and Poly(A) RNAs
摘要: The ability to bind strongly to complementary nucleic acid sequences, invade complex nucleic acid structures, and resist degradation by cellular enzymes has made peptide nucleic acid (PNA) oligomers as very useful hybridization probes in molecular diagnosis. For such applications, the PNA oligomers have to be labeled with appropriate reporters as they lack intrinsic labels that can be used in biophysical assays. Although solid-phase synthesis is commonly used to attach reporters onto PNA, development of milder and modular labeling methods will provide access to PNA oligomers labeled with a wider range of biophysical tags. Here, we describe the establishment of a postsynthetic modification strategy based on bioorthogonal chemical reactions in functionalizing PNA oligomers in solution with a variety of tags. A toolbox composed of alkyne- and azide-modified monomers were site-specifically incorporated into PNA oligomers and postsynthetically click-functionalized with various tags, ranging from sugar, amino acid, biotin, to fluorophores, by using copper(I)-catalyzed azide?alkyne cycloaddition, strain-promoted azide?alkyne cycloaddition, and Staudinger ligation reactions. As a proof of utility of this method, fluorescent PNA hybridization probes were developed and used in imaging human telomeres in chromosomes and poly(A) RNAs in cells. Taken together, this simple approach of generating a wide range of functional PNA oligomers will expand the use of PNA in molecular diagnosis.
关键词: peptide nucleic acid,bioorthogonal chemical reactions,molecular diagnosis,poly(A) RNAs,human telomeres,PNA,click chemistry
更新于2025-09-09 09:28:46
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PEGylated Fluorescent Nanoparticles from One-Pot Atom Transfer Radical Polymerization and “Click Chemistry”
摘要: The preparation of PEGylated fluorescent nanoparticles (NPs) based on atom transfer radical polymerization (ATRP) and “click chemistry” in one-pot synthesis is presented. First, poly(p-chloromethyl styrene-alt-N-propargylmaleimide) (P(CMS-alt-NPM)) copolymer was prepared via reversible addition-fragmentation chain transfer (RAFT) polymerization. Subsequently, the azido-containing fluorene-based polymer, poly[(9,9-dihexylfluorene)-alt-(9,9-bis-(6-azidohexyl)fluorene)] (PFC6N3), was synthesized via Suzuki coupling polymerization, followed by azidation. Finally, the PEGylated fluorescent NPs were prepared via simultaneous intermolecular “click” cross-linking between P(CMS-alt-NPM) and PFC6N3 and the ATRP of poly(ethylene glycol) methyl ether methacrylate (PEGMMA) using P(CMS-alt-NPM) as the macroinitiator. The low cytotoxicity of the PEGylated fluorescent NPs was revealed by incubation with KB cells, a cell line derived from carcinoma of the nasopharynx, in an in vitro experiment. The biocompatible PEGylated fluorescent NPs were further used as a labeling agent for KB cells.
关键词: ATRP,fluorescent nanoparticles,click chemistry,one-pot synthesis,PEGylated,cellular imaging
更新于2025-09-09 09:28:46
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A Versatile Dynamic Light Scattering Strategy for the Sensitive Detection of Plant MicroRNAs Based‐on Click Chemistry‐Amplified Aggregation of Gold Nanoparticles
摘要: Plant microRNAs (miRNAs) are naturally 2′-O-methylated at the 3′-terminal, so they cannot be efficiently detected by traditional target-triggered polymerization reactions. Herein, a simple but robust enzyme-free sensing strategy is developed for plant miRNA analysis by using dynamic light scattering (DLS) technique to monitor the click chemical ligation-amplified crosslinking of gold nanoparticles (AuNPs). Combining the enzyme-free cycling chemical ligation-mediated signal amplification, and the intrinsic outstanding ability of DLS for discriminating the extremely low level of particle aggregation in a large pool of monodisperse AuNPs, high sensitivity is achieved and as low as 78.6 fM plant miRNA can be easily detected.
关键词: crosslinking,click chemistry,plant microRNA,dynamic light scattering (DLS),gold nanoparticles
更新于2025-09-09 09:28:46
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Multichromophoric sugar for fluorescence photoswitching
摘要: A multichromophoric glucopyranoside 2 bearing three dicyanomethylenepyran (DCM) fluorophores and one diarylethene (DAE) photochrome has been prepared by Cu(I)-catalyzed alkyne–azide cycloaddition reaction. The fluorescence of 2 was switched off upon UV irradiation, in proportion with the open to closed form (OF to CF) conversion extent of the DAE moiety. A nearly 100% F?rster-type resonance energy transfer (FRET) from all three DCM moieties to a single DAE (in its CF) moiety was achieved. Upon visible irradiation, the initial fluorescence intensity was recovered. The observed photoswiching is reversible, with excellent photo resistance.
关键词: energy transfer,monosaccharide,photochromism,click chemistry,fluorophore
更新于2025-09-04 15:30:14