Product Focus: 4-Thio-UTP: Photochemical Cross-Linking and Affinity Selection

The base-modified UTP analog 4-Thio-UTP can be efficiently incorporated into RNA by in vitro transcription[1]. The resulting thiolated RNA is suitable for photochemical cross-linking for characterization of RNA-protein interactions in ribonucleoprotein (RNP) complexes[2,3] or for affinity RNA selection experiments[4,5]. Further, 4-Thio-UTP was shown to be involved in the stimulation of human P2Y2 and P2Y4 receptors[6].

Applications of 4-Thio-UTP that were recently reported in the literature:

Field of use

Affinity RNA selection experiment

Application

Development of an alternative means to isolate purer nascent RNA fractions. 4-Thio-UTP labeled nascent RNA was biotinylated and specifically enriched from non-labeled RNA following capture by streptavidin coated beads.

Reference

Davidson et al. (2012) Co-transcriptional degradation of aberrant pre-mRNA by Xrn2. The EMBO Journal 31:2566.

Field of use

Photochemical cross-linking

Application

In vitro transcription of wild-type and mutant IBV (Infectious Bronchitis Virus) pseudoknot RNAs in the presence of 4-Thio-UTP, incubation of IBV pseudoknot RNA with cell extracts of human lung fibroblast IMR90 and subsequent UV irradiation to directly crosslink interacting proteins to 4-Thio-UTP incorporated IBV pseudoknot RNA.

Reference

Kwak et al. (2011) Annexin A2 binds RNA and reduces the frameshifting efficiency of Infectious Bronchitis Virus. PLoS One 6(8):e24067.

Selected References

  1. [Nilsen et al. (2012) In vitro transcription of labeled RNA: Synthesis, capping, and substitution. Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory Press, 1181.
  2. Temiakov et al. (2003) Probing the organization of transcription complexes using photoreactive 4-thio-substituted analogs of uracil and thymidine. Methods in Enzymology 371:133.
  3. Hanna (1989) Photoaffinity cross-linking methods for studying RNA-protein interactions. Methods in Enzymology 180:383.
  4. Davidson et al. (2012) Co-transcriptional degradation of aberrant pre-mRNA by Xrn2. The EMBO Journal 31:2566.
  5. Kenzelmann et al. (2007) Microarray analysis of newly synthesized RNA in cells and animals. PNAS 104(15):6164.
  6. Brunschweiger et al. (2006) P2 receptors activated by uracil nucleotides – an update. Current Medicinal Chemistry 13:289.

Where can I find more information about Jena Bioscience?

Visit the manufacturer page at www.stratech.co.uk/jena_bioscience, email info@stratech.co.uk or call +44 (0) 1638 782600.

Stratech Scientific is a distributor of high quality, competitively priced, reliable products for research laboratories throughout the UK and Europe. Please contact us to find out which ranges we can supply in your country.

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