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Method of generating branched and multi-chain nucleic acid switches for ligand detection

  • US 7,829,287 B2
  • Filed: 03/10/2009
  • Issued: 11/09/2010
  • Est. Priority Date: 08/03/2004
  • Status: Active Grant
First Claim
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1. A method to generate a multichain nucleic acid switch adapted to switch from a first conformation to a second conformation upon ligand binding, said switch comprising:

  • a probe strand P comprising a ligand binding domain;

    a switching framework comprising a cover strand (C);

    a tether comprising a base-pair stem that holds P and C together while the switch changes between the first and second conformations, wherein P and C are on different nucleic acid chains; and

    a signaling apparatus comprising a combination of signaling entities, said method comprising the following steps;

    (i) altering two nucleic acid sequences (a) and (b) to introduce mismatches between P and C;

    (ii) measuring the signal for the two nucleic acid sequences (a) and (b), thereby determining an equilibrium constant, K1 for the switch from the first conformation to the second conformation for each of the two sequences (a) and (b);

    (iii) adjusting the equilibrium constant, K1 between 0.1-0.01 based on K1 for sequences (a) and (b); and

    (iv) generating the multichain nucleic acid switch having the equilibrium constant, K1 between 0.1-0.01.

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