| Versatile phosphoramidation reactions for nucleic acid conjugations with peptides, proteins, chromophores, and biotin derivatives. | |
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MedLine Citation:
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PMID: 20690641 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Chemical conjugations of nucleic acids with macromolecules or small molecules are common approaches to study nucleic acids in chemistry and biology and to exploit nucleic acids for medical applications. The conjugation of nucleic acids such as oligonucleotides with peptides is especially useful to circumvent cell delivery and specificity problems of oligonucleotides as therapeutic agents. However, current approaches are limited and inefficient in their ability to afford peptide-oligonucleotide conjugates (POCs). Here, we report an effective and reproducible approach to prepare POCs and other nucleic acid conjugates based on a newly developed nucleic acid phosphoramidation method. The development of a new nucleic acid phosphoramidation reaction was achieved by our successful synthesis of a novel amine-containing biotin derivative used to systematically optimize the reactions. The improved phosphoramidation reactions dramatically increased yields of nucleic acid-biotin conjugates up to 80% after 3 h reaction. Any nucleic acids with a terminal phosphate group are suitable reactants in phosphoramidation reactions to conjugate with amine-containing molecules such as biotin and fluorescein derivatives, proteins, and, most importantly, peptides to enable the synthesis of POCs for therapeutic applications. Polymerase chain reactions (PCRs) to study incorporation of biotin or fluorescein-tagged DNA primers into the reaction products demonstrated that appropriate controls of nucleic acid phosphoramidation reactions incur minimum adverse effects on inherited base-pairing characteristics of nucleotides in nucleic acids. The phosphoramidation approach preserves the integrity of hybridization specificity in nucleic acids when preparing POCs. By retaining integrity of the nucleic acids, their effectiveness as therapeutic reagents for gene silencing, gene therapy, and RNA interference is ensured. The potential for POC use was demonstrated by two-step phosphoramidation reactions to successfully synthesize nucleic acid-tetraglycine conjugates. In addition, phosphoramidation reactions provided a facile approach to prepare nucleic acid-BSA conjugates with good yields. In summary, the new approach to phosphoramidation reactions offers a universal method to prepare POCs and other nucleic acid conjugates with high yields in aqueous solutions. The methods can be easily adapted to typical chemistry or biology laboratory setups which will expedite the applications of POCs for basic research and medicine. |
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Authors:
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Tzu-Pin Wang; Yi-Jang Chiou; Yi Chen; Eng-Chi Wang; Long-Chih Hwang; Bing-Hung Chen; Yen-Hsu Chen; Chun-Han Ko |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Bioconjugate chemistry Volume: 21 ISSN: 1520-4812 ISO Abbreviation: Bioconjug. Chem. Publication Date: 2010 Sep |
Date Detail:
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Created Date: 2010-09-15 Completed Date: 2010-12-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9010319 Medline TA: Bioconjug Chem Country: United States |
Other Details:
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Languages: eng Pagination: 1642-55 Citation Subset: IM |
Affiliation:
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Department of Medicinal and Applied Chemistry, Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amines
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chemistry Biotin / analogs & derivatives* Fluoresceins / chemistry Nucleic Acids / chemistry* Organophosphorus Compounds / chemistry* Peptide Nucleic Acids / chemical synthesis*, chemistry Peptides / chemistry* Phosphates / chemistry Polymerase Chain Reaction |
| Chemical | |
Reg. No./Substance:
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0/Amines; 0/Fluoresceins; 0/Nucleic Acids; 0/Organophosphorus Compounds; 0/Peptide Nucleic Acids; 0/Peptides; 0/Phosphates; 0/phosphoramidite; 58-85-5/Biotin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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