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Single-molecule imaging of an in vitro-evolved RNA aptamer reveals homogeneous ligand binding kinetics

Journal Article


Abstract


  • Many studies of RNA folding and catalysis have revealed conformational heterogeneity, metastable folding intermediates, and long-lived states with distinct catalytic activities. We have developed a single-molecule imaging approach for investigating the functional heterogeneity of in vitro-evolved RNA aptamers. Monitoring the association of fluorescently labeled ligands with individual RNA aptamer molecules has allowed us to record binding events over the course of multiple days, thus providing sufficient statistics to quantitatively define the kinetic properties at the single-molecule level. The ligand binding kinetics of the highly optimized RNA aptamer studied here displays a remarkable degree of uniformity and lack of memory. Such homogeneous behavior is quite different from the heterogeneity seen in previous single-molecule studies of naturally derived RNA and protein enzymes. The single-molecule methods we describe may be of use in analyzing the distribution of functional molecules in heterogeneous evolving populations or even in unselected samples of random sequences.

Authors


Publication Date


  • 2009

Citation


  • Elenko, M. P., Szostak, J. W. & van Oijen, A. M. (2009). Single-molecule imaging of an in vitro-evolved RNA aptamer reveals homogeneous ligand binding kinetics. Journal of the American Chemical Society, 131 (29), 9866-9867.

Scopus Eid


  • 2-s2.0-67651208284

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2198

Has Global Citation Frequency


Number Of Pages


  • 1

Start Page


  • 9866

End Page


  • 9867

Volume


  • 131

Issue


  • 29

Place Of Publication


  • United States

Abstract


  • Many studies of RNA folding and catalysis have revealed conformational heterogeneity, metastable folding intermediates, and long-lived states with distinct catalytic activities. We have developed a single-molecule imaging approach for investigating the functional heterogeneity of in vitro-evolved RNA aptamers. Monitoring the association of fluorescently labeled ligands with individual RNA aptamer molecules has allowed us to record binding events over the course of multiple days, thus providing sufficient statistics to quantitatively define the kinetic properties at the single-molecule level. The ligand binding kinetics of the highly optimized RNA aptamer studied here displays a remarkable degree of uniformity and lack of memory. Such homogeneous behavior is quite different from the heterogeneity seen in previous single-molecule studies of naturally derived RNA and protein enzymes. The single-molecule methods we describe may be of use in analyzing the distribution of functional molecules in heterogeneous evolving populations or even in unselected samples of random sequences.

Authors


Publication Date


  • 2009

Citation


  • Elenko, M. P., Szostak, J. W. & van Oijen, A. M. (2009). Single-molecule imaging of an in vitro-evolved RNA aptamer reveals homogeneous ligand binding kinetics. Journal of the American Chemical Society, 131 (29), 9866-9867.

Scopus Eid


  • 2-s2.0-67651208284

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/2198

Has Global Citation Frequency


Number Of Pages


  • 1

Start Page


  • 9866

End Page


  • 9867

Volume


  • 131

Issue


  • 29

Place Of Publication


  • United States