RNA complexes methods of their production and sensors and analytical methods involving same

A method of determining the presence of an analyte through the formation a self-assembled RNA complex, said method comprising the steps:

• (a) placing in solution at least three monomeric units of an RNA molecule, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, at least one of said monomeric units comprising an aptamer adapted to react with said analyte;
• and
• (b) allowing said at least one of said monomeric units comprising an aptamer to react with said analyte, and such that, in the presence of said analyte, said at least three monomeric units self-assemble by respective first binding sites interacting with first binding sites of respective adjacent monomeric units, and respective second binding sites interacting with second binding sites of respective adjacent monomeric units to form pairs of cognate interactions and so as to form said RNA complex in a circular closed complex;
• and
• (c) determining the presence or absence of said complex.

A sensor adapted to bind an analyte, said sensor comprising:

• (a) a vessel containing a solution of at least three monomeric units of an RNA molecule, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, at least one of said monomeric units comprising an aptamer adapted to react with said analyte such that said at least three monomeric units are adapted to self-assemble in the presence of said analyte by respective first binding sites interacting with first binding sites of respective adjacent monomeric units, and respective second binding sites interacting with second binding sites of respective adjacent monomeric units, to form pairs of cognate interactions and so as to form said RNA complex in a circular closed complex;
• and
• (b) means to detect the presence or absence of said complex.
• Method of Determining Presence of Two or More Analytes by Formation of an RNA Complex with at least one of the Monomers having an Aptamer reacting with the Analyte

A method of determining the presence of at least two analytes in solution through the formation respective self-assembled RNA complexes, said method comprising the steps:

• (a) placing in contact with said solution the surface of two silicon cantilevers bound to respective monomeric units of an RNA molecule of a respective first and second type, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, and said monomeric unit of said first type comprising an aptamer adapted to react with the first of said two analytes, and said monomeric unit of said second type comprising an aptamer adapted to react with the second of said two analytes;
• (b) said solution containing additional monomeric units of an RNA molecule of a third and fourth type, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, such that monomeric units of said first, third and fourth types self-assemble by respective first binding sites interacting with first binding sites of respective adjacent monomeric units, and respective second binding sites interacting with second binding sites of respective adjacent monomeric units to form pairs of cognate interactions and so as to form said RNA complex in a circular closed complex on said respective silicon cantilever surface should said first analyte be present in said solution;
• and such that monomeric units of said second, third and fourth types self-assemble by respective first binding sites interacting with first binding sites of respective adjacent monomeric units, and respective second binding sites interacting with second binding sites of respective adjacent monomeric units to form pairs of cognate interactions and so as to form said RNA complex in a circular closed complex on said respective silicon cantilever surface should said second analyte be present in said solution;
• and
• (b) determining the presence or absence of said complexes on said respective silicon cantilever surfaces.
• Cantilever Detection Apparatus having Cantilevers with Analyte-Specific Aptamer-Bearing Monomers

A method of identifying an RNA molecule capable of forming a self-assembled RNA complexes with RNA molecules of two known types, said method comprising the steps:

• (a) placing in solution:
• (i) monomeric units of an RNA molecule of a respective a first and second type, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, such that said first and second types are adapted to bind to one another and;
• (ii) a plurality of monomeric units of an RNA molecule of a third type, each said monomeric unit comprising RNA having a first helical domain having a first and second binding site, said first helical domain connected to a second helical domain having a first and second binding site, said first and second binding sites of said third type first helical domain adapted to bind to respective first and second binding sites of said first type first helical domain, and said first binding site of said third type second helical domain adapted to bind to respective first binding site of said second type second helical domain, and said second binding site of said third type second helical domain being randomized, such that only some of said monomeric units of an RNA molecule of said third type self-assemble with said first and second types by respective adjacent monomeric units forming pairs of cognate interactions and so as to form said RNA complex in a circular closed complex;
• (b) separating from said solution said RNA complex so formed;
• and
• (c) dissociating said RNA molecule of said third type from said RNA complex so as to isolate an RNA molecule of said third type.