Single stranded RNA-oligonucleotides, phosphorothioate (PTO-) protected
Scientific context
Toll-like receptors (TLRs) are fundamental for the innate immune response. Two of these orphan receptors TLR7/TLR8 have been identified to act as sensors for single stranded RNA. Whereas doublestranded ribonucleic acid (dsRNA) was already known as a danger signal associated with viral infection and stimulates innate immune cells, the immunostimulatory potential of single-stranded RNA (ssRNA) is poorly understood and respective receptors were unknown. Only recently members of the TLR family were identified to play an important role in the sensing of ssRNAs. Activation of TLRs leads to the generation of an adoptive immune response resulting in the eradication of pathogens. Guanosine (G)-and uridine (U)-rich ssRNA oligonucleotides derived from human immunodeficiency virus-1 (HIV-1) stimulate dendritic cells (DCs) and macrophages to secrete interferon-α and proinflammatory, as well as regulatory, cytokines.
Beside a research group at the TU Munich two other groups at the Imperial Cancer Research (ICRF) in London and the other at the Howard Hughes Medical Institute at Yale University in New Haven also reported on this phenomenon. Researchers in the group of Stefan Bauer at the TU Munich stated that this discovery has an interesting potential for the development of novel methods of immunization for vaccine design using RNA as adjuvant.
Mediators of the innate immune system for the sensing of bacterial pathogens such as CpG-rich motifs have already entered clinical trials.
Products
IBAs GeneTAGnology division has a long lasting experience not only in DNA-oligonucleotide synthesis, but also in the synthesis of high quality single-stranded RNA molecules. Products of our GeneTAGnology division have been used amongst others by a research group at the TU munich to identify members of the Toll-like receptor (TLR) family as sensors for single stranded RNA.
IBA Quality
IBA ssRNA as well as our other RNA products are manufactured in state-of-the art facilities. Due to the phosphorothioate modifications the stability of this kind of ssRNA-oligonucleotides is increased significantly. Quality is checked routinely according to our stringent quality standards. Special care is taken to use only highest quality raw materials. IBAs ssRNA products are especially recognized for their superior performance in cell culture assays looking for an immune response monitoring avoiding any kind of endotoxin-related unspecific effects.
All modifications which are available for DNA are available for RNA Phosphorothioates too
Tools for enzymatic DNA and RNA modifications
Modified triphosphates
Scientific context
Transcription analysis is an important tool not only in basic molecular biology but also in the investigation of the molecular mechanisms of a variety of diseases. Examples amongst these are certain types of anemia or xeroderma pigmentosum. In this context labeled nucleoside triphophates (NTP) are used widely to monitor RNA synthesis in vitro. Other application using modified triphosphates are the systematic evolution of ligands by exponential enrichment, a method to screen for functional RNA oligomers either working as catalytically active RNA or as a specific class of target directed RNA oligomers called aptamers.
The potential of standard in vitro transcription reactions can be dramatically expanded, if chemically synthesized low-mol-weight compounds are used as building blocks in combination with standard nucleotide 5' triphosphates (NTPs). Clearly, chemically synthesized, modified NTPs are inserted at internal sites. The combination with phosphorothioate linkages for detection has been developed into a powerful high-throughput method to study site-specific interference of modifications with RNA function.
OLSON, DB, SAYERS, JR, ECKSTEIN, F, 1993: Methods ENZYMOL 217: 189 - 217.
Labeling of PCR products via incorporation of Biotin-dUTP
*see price list for details
Dinucleotides
5'-Modified ApG can be used in vitro transcription experiments. It is efficiently incorporated at the 5'-end. By using this compound it is easy to either introduce fluorescent markers to make transcripts visible, or non-radioactive markers like biotin or digoxigenin to isolate transcription products. BiotindUTP is used for labeling of PCR products.
Toll-like receptors (TLRs) are fundamental for the innate immune response. Two of these orphan receptors TLR7/TLR8 have been identified to act as sensors for single stranded RNA. Whereas doublestranded ribonucleic acid (dsRNA) was already known as a danger signal associated with viral infection and stimulates innate immune cells, the immunostimulatory potential of single-stranded RNA (ssRNA) is poorly understood and respective receptors were unknown. Only recently members of the TLR family were identified to play an important role in the sensing of ssRNAs. Activation of TLRs leads to the generation of an adoptive immune response resulting in the eradication of pathogens. Guanosine (G)-and uridine (U)-rich ssRNA oligonucleotides derived from human immunodeficiency virus-1 (HIV-1) stimulate dendritic cells (DCs) and macrophages to secrete interferon-α and proinflammatory, as well as regulatory, cytokines.