In vivo identification of ribonucleoprotein–RNA interactions
2006 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 103, no 5, 1557-1562 p.Article in journal (Refereed) Published
To understand the role of RNA-binding proteins (RBPs) in the regulation of gene expression, methods are needed for the in vivo identification of RNA–protein interactions. We have developed the peptide nucleic acid (PNA)-assisted identification of RBP technology to enable the identification of proteins that complex with a target RNA in vivo. Specific regions of the 3ˈ and 5ˈ UTRs of ankylosis mRNA were targeted by antisense PNAs transported into cortical neurons by the cell-penetrating peptide transportan 10. An array of proteins was isolated in complex with or near the targeted regions of the ankylosis mRNA through UV-induced crosslinking of the annealed PNA–RNA–RBP complex. The first evidence for pharmacological modulation of these specific protein–RNA associations was observed. These data show that the PNA-assisted identification of the RBP technique is a reliable method to rapidly identify proteins interacting in vivo with the target RNA.
Place, publisher, year, edition, pages
2006. Vol. 103, no 5, 1557-1562 p.
Microbiology in the medical area
IdentifiersURN: urn:nbn:se:su:diva-39089DOI: 10.1073/pnas.0510611103OAI: oai:DiVA.org:su-39089DiVA: diva2:318348
Jennifer Zielinski and Kalle Kilk contributed equally to this work.2010-05-072010-05-072015-04-21Bibliographically approved