Preparation of Oligodeoxynucleotide 5′-Triphosphates Using Solid Support Approach

Lebedev, A.; Koukhareva, Inna; Beck, Terry; Vaghefi, Morteza M.

doi:10.1081/ncn-100002565

Cited by 24 publications

(16 citation statements)

References 3 publications

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“…[14] Then, the oxidation reaction was performed with elemental sulfur in DMF, [15] and the cyclic triphosphate was opened by aqueous buffered treatment before ON deprotection under basic conditions. [16] Owing to this moderate yield, we further investigated a more effective method for the formation of the αthio TP bond. These data were consistent with the work reported on the synthesis of 5Ј-TP ONs on a solid support by using the Ludwig and Eckstein method.…”

Section: Resultsmentioning

confidence: 99%

Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates

et al. 2014

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A robust solid‐phase synthesis was developed to obtain original oligonucleotides (ONs) functionalized at their 5′ end with modified triphosphate (TP) moieties, in which a nonbridging oxygen atom of the α phosphorus atom was replaced by a sulfur atom and the labile P–O–P linkage was changed into a methylene bridge between the β and γ phosphorus atoms. The efficient method is based on solid‐supported ON assembly followed by 5′‐H‐phosphonylation, oxidation to the thiophosphate subsequently activated as a phosphoanhydride with diphenyl phosphoryl chloride, then nucleophilic substitution with the alkylammonium salt of pyrophosphate or its β,γ‐methylene analogue. After deprotection and release from the solid support under basic conditions, 5′‐(α‐P‐thio)TP and 5′‐(α‐P‐thio)(β,γ‐methylene)TP oligonucleotides were obtained in satisfactory yields, and they were isolated with high purity. These hydrolysis‐resistant 5′‐TP ONs will be useful in biological research to elucidate the mechanism of enzymes involved in mRNA processing and maturation.

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Section: Resultsmentioning

confidence: 99%

Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates

et al. 2014

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“…A number of modified nucleoside 5′‐triphosphates have been prepared by using 2‐chloro‐4 H ‐1,3,2‐benzodioxaphosphorin‐4‐one . This reagent was also used in the synthesis of oligonucleotide 5′‐triphosphates on a solid support …”

Section: The Use Of Compounds Containing a P−cl Bondmentioning

confidence: 99%

How To Form a Phosphate Anhydride Linkage in Nucleotide Derivatives

Sherstyuk

Abramova

2015

ChemBioChem

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The fundamental roles of nucleoside triphosphates and nucleotide cofactors such as NAD(+) in biochemistry are well known. In recent decades, continuing research has revealed the key role of 5'-capped RNA and 5',5'-dinucleoside polyphosphates in the regulation of vitally important physiological processes. Last but not least, the commercial potential of nucleoside triphosphate synthesis can hardly be overestimated. Nevertheless, despite decades of investigation and the obvious topicality of the research on the chemical synthesis of the nucleotide compounds containing phosphate anhydride linkages, none of the existing procedures can be considered an up-to-date "gold standard". However, there are a number of fruitful synthetic approaches to forming phosphate anhydride linkages in satisfactory yield. These are summarized in this concise review, organized by the type of active phosphorous intermediate and reagents used.

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“…Despite the existence of an arsenal of methods for the synthesis of nucleoside triphosphates (Burgess and Cook, ), to date, there is still no efficient and universal method for the enzyme‐free chemical synthesis of 5′‐triphosphate DNA and RNA. The few approaches describing their synthesis on solid support (Brownlee et al, ; Olsen et al, ; Burgess and Cook, ; Lebedev et al, ) are all accompanied by flaws such as dependence on oligonucleotide length and sequence, difficult separation procedures resulting from low conversions, and poor yields. These approaches are also somewhat limited to the small‐scale production of triphosphate oligonucleotides, and do not demonstrate compatibility with possible chemical modifications within the oligonucleotide backbone.…”

Section: Commentarymentioning

confidence: 99%

Solid‐Phase Chemical Synthesis of 5′‐Triphosphate DNA, RNA, and Chemically Modified Oligonucleotides

Zlatev

Manoharan

Vasseur

et al. 2012

CP Nucleic Acid Chemistry

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A chemical method for the solid-phase synthesis of 5'-triphosphate oligonucleotides is described. The full-length oligonucleotides are first constructed using standard solid-phase DNA/RNA synthesis, and then efficient implementation of a sequential 4-steps synthetic procedure, executed either manually or in a fully automated fashion, affords the corresponding solid-supported 5'-triphosphate oligonucleotides. Using this synthetic procedure, the full-length 5'-hydroxyl oligonucleotides are initially transformed into the corresponding 5'-H-phosphonate mono esters, subsequently oxidized in the presence of imidazole to the activated 5'-phosphorimidazolidates, and finally reacted with pyrophosphate on the solid support. The method uses safe, stable, and inexpensive reagents, and the process is scalable and readily applicable to automated synthesis compatible with the current commercially available DNA/RNA synthesizers. After cleavage from the solid support and deprotection, a range of DNA, RNA, and chemically modified 5'-triphosphate oligonucleotides are obtained in a convenient and efficient manner and isolated in good yields after HPLC purification.

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Preparation of Oligodeoxynucleotide 5′-Triphosphates Using Solid Support Approach

Cited by 24 publications

References 3 publications

Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates

Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates

How To Form a Phosphate Anhydride Linkage in Nucleotide Derivatives

Solid‐Phase Chemical Synthesis of 5′‐Triphosphate DNA, RNA, and Chemically Modified Oligonucleotides

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