PMID: 11345441May 10, 2001Paper

Streptavidin aptamers: affinity tags for the study of RNAs and ribonucleoproteins

RNA
Chatchawan Srisawat, D R Engelke

Abstract

RNA affinity tags would be very useful for the study of RNAs and ribonucleoproteins (RNPs) as a means for rapid detection, immobilization, and purification. To develop a new affinity tag, streptavidin-binding RNA ligands, termed "aptamers," were identified from a random RNA library using in vitro selection. Individual aptamers were classified into two groups based on common sequences, and representative members of the groups had sufficiently low dissociation constants to suggest they would be useful affinity tools. Binding of the aptamers to streptavidin was blocked by presaturation of the streptavidin with biotin, and biotin could be used to dissociate RNA/streptavidin complexes. To investigate the practicality of using the aptamer as an affinity tag, one of the higher affinity aptamers was inserted into RPR1 RNA, the large RNA subunit of RNase P. The aptamer-tagged RNase P could be specifically isolated using commercially available streptavidin-agarose and recovered in a catalytically active form when biotin was used as an eluting agent under mild conditions. The aptamer tag was also used to demonstrate that RNase P exists in a monomeric form, and is not tightly associated with RNase MRP, a closely related ribonucleoprotein e...Continue Reading

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Citations

Jun 27, 2012·Analytical and Bioanalytical Chemistry·M SvobodováC K O' Sullivan
Sep 24, 2013·Applied Microbiology and Biotechnology·Christopher M DundasSheldon Park
Jun 11, 2002·Methods : a Companion to Methods in Enzymology·Chatchawan Srisawat, David R Engelke
Jun 11, 2002·Methods : a Companion to Methods in Enzymology·Somashe NiranjanakumariMariano A Garcia-Blanco
Nov 15, 2002·Methods : a Companion to Methods in Enzymology·Melissa S Jurica, Melissa J Moore
Oct 31, 2002·RNA·Chatchawan SrisawatDavid R Engelke
Jul 23, 2013·Nature Methods·Wenjiao SongSamie R Jaffrey
Jun 15, 2007·Nature Methods·Luc Ponchon, Frédéric Dardel
Aug 21, 2008·Proceedings of the National Academy of Sciences of the United States of America·Daniel J CoughlinDavid R Engelke
Jun 23, 2009·Proceedings of the National Academy of Sciences of the United States of America·Falk ButterMatthias Mann
Aug 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Ho TsoiHo Yin Edwin Chan
Mar 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Ho Young LeeJennifer A Doudna
May 10, 2002·Nucleic Acids Research·Abdessamad Tahiri-AlaouiWilliam James
Apr 22, 2009·Nucleic Acids Research·Daiying Xu, Hua Shi
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