Journal ArticleDOI
An efficient method for the isolation and purification of oligoribonucleotides
Brian Sproat,Francesco Colonna,Bashar Mullah,Dean Tsou,Alex Andrus,Arnold Hampel,Ravi Vinayak +6 more
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TLDR
An efficient anion-exchange HPLC method has been developed for the purification of chemically synthesized RNA and the resulting product precipitated directly by the addition of 1-propanol and a new activator, 5-ethylthio-1H-tetrazole significantly enhances the synthesis quality and yield of oligoribonucleotides.Abstract:
Problems associated with the use of tetrabutylammonium fluoride like incomplete desilylation and removal of the tetrabutylammonium salts during large scale syntheses of oligoribonucleotides (RNA) have been eliminated by the use of triethylamine trihydrofluoride and precipitation of the RNA with 1-butanol. An efficient anion-exchange HPLC method has been developed for the purification of chemically synthesized RNA and the resulting product precipitated directly by the addition of 1-propanol. A new activator, 5-ethylthio-1H-tetrazole significantly enhances the synthesis quality and yield of oligoribonucleotides. RNA synthesized using these improvements has been shown to be biologically active by a comparative ribozyme-substrate assay.read more
Citations
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Journal ArticleDOI
NMR investigation of RNA structure
Journal ArticleDOI
Oligo- and poly-nucleotides: 50 years of chemical synthesis
TL;DR: It is fifty years since the first chemical synthesis of a dinucleoside phosphate and a din nucleotide with natural 3'-->5'-internucleotide linkages was reported.
Journal ArticleDOI
Tertiary structure formation in the hairpin ribozyme monitored by fluorescence resonance energy transfer
TL;DR: It is demonstrated directly that mutations and modifications which inhibit a step following substrate binding are not necessarily involved in catalysis, including specific structural transitions of the catalytic cycle.
Journal ArticleDOI
Transfer RNA recognition by aminoacyl-tRNA synthetases.
TL;DR: The structural information available for synthetases is reviewed, the insights into specific recognition gained from classical and atomic group mutagenesis experiments performed with tRNAs, tRNA fragments, and small RNAs mimicking portions of tRNA are explored.
Journal ArticleDOI
Inter-domain cross-linking and molecular modelling of the hairpin ribozyme.
David J. Earnshaw,Benoît Masquida,Sabine Müller,Snorri Th. Sigurdsson,Fritz Eckstein,Eric Westhof,Michael J. Gait +6 more
TL;DR: The hairpin ribozyme is a small catalytic RNA composed of two helical domains containing a small and a large internal loop and constitutes a valuable paradigm for the study of RNA structure and catalysis as mentioned in this paper.
References
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Journal ArticleDOI
Antisense oligonucleotides: a new therapeutic principle
Eugen Uhlmann,Anusch Peyman +1 more
Journal ArticleDOI
Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates
TL;DR: A method is described to synthesize small RNAs of defined length and sequence using T7 RNA polymerase and templates of synthetic DNA which contain the T7 promoter to increase the variety of RNAs which can be made.
Book
Oligonucleotides and analogues : a practical approach
TL;DR: The present work presents modern machine-aided methods of oligodeoxyribonucleotide synthesis with a focus on the synthesis of oligonucleotides attached to intercalators, photoreactive agents and cleavage agents.
Journal ArticleDOI