The solid phase synthesis of RNA fragments is now a widespread technique in Molecular Biology. RNA oligonucleotides are becoming increasingly important in structural studies where they can be used as probes in Northern blotting or for in situ hybridization. In regulation, RNA oligonucleotides have been used in an antisense mode. The role of catalytic RNA (ribozymes and most importantly siRNA) as putative antisense therapeutics has sparked considerable interest in synthetic RNA.
Although on the whole it’s similar to DNA synthesis the additional 2’-OH reactive group of RNA introduces considerable complexity to the synthesis and requires an additional protective group. The use of the 2’-O-TBDMS protecting group has now become widely accepted as the best compromise for RNA synthesis of oligonucleotides up to 80 bases long.
Standard RNA oligonucleotides
Recent discoveries regarding catalytic activity of some types of RNA molecules (e.g. the «ribozymes»), the role of cellular RNA as target for antisense therapeutics, the possible therapeutic role for modified RNAs, and the recent discovery of siRNA underscore the growing importance of synthetic RNA in basic biomedical research.
The increased demand for synthetic RNA oligonucleotides led to the development of specially modified reagents and protocols for the routine synthesis of high-quality RNA oligonucleotides. It is now possible to synthesize an RNA molecule up to 80 bases. The automated synthesis of RNA oligonucleotides is technically a rather more difficult task than the synthesis of DNA oligonucleotides, due primarily to the presence of the 2’-OH group and due to the susceptibility of the RNA product to degradation.
Eurogentec has met the challenge of RNA oligonucleotide synthesis by using a combination of base-labile protecting groups and the 2’-O-TBDMS protection system, which offers the best approach to RNA synthesis currently available. This, together with an exclusive Eurogentec protocol for automated synthesis, results in synthesis of RNA oligonucleotides of excellent purity, stability and biological activity. We also have the possibility to synthesize DNA/RNA chimeric oligos. RNA oligos can be labelled with fluorescent dyes or with Biotin. RNA oligos are delivered lyophilized under RNase-free conditions to ensure the best stability upon transportation.
siRNA synthesis
Discovered in 1998, RNA interference (RNAi) represents the major technological advance in Molecular Biology since the discovery of PCR. RNA interference is a mechanism of gene silencing at the mRNA level. It offers a quick, easy, efficient and broadly applicable approach for gene knock-out. At least, a technology that will allow you to unravel the function of your gene of interest! The easiest and most efficient way to achieve RNAi is to use synthetic small-interfering RNA (siRNA). siRNA are duplexes of short mixed oligonucleotides. These are typically made of 19 RNAs nucleotides with symmetric 2 nucleotides 3’ overhangs.
Virtually, any gene can be targeted by RNA interference! RNAi methods based on synthetic siRNA require minimal time and labour; therefore, mammalian gene knockdown by RNAi as become very popular during the last years.
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