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SELEX AND PHOTOSELEX

  • US 20090098549A1
  • Filed: 07/17/2008
  • Published: 04/16/2009
  • Est. Priority Date: 07/17/2007
  • Status: Active Grant
First Claim
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1. A method for identifying a photoaptamer, the method comprising:

  • a) preparing a candidate mixture of nucleic acids, wherein each nucleic acid comprises;

    i) at least one non-photoreactive placeholding pyrimidine; and

    ii) at least one modified pyrimidine independently selected from the group consisting of the modified pyrimidines shown in FIG. 2;

    b) contacting the candidate mixture with a target, wherein nucleic acids having an increased affinity to the target relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;

    c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture;

    d) amplifying the increased affinity nucleic acids to yield a nucleic acid ligand-enriched mixture of nucleic acids;

    e) repeating (b)-(d) as desired;

    f) producing from said nucleic acid ligand-enriched mixture of nucleic acids a candidate photoaptamer or a mixture of candidate photoaptamers by replacing in each nucleic acid of the ligand-enriched mixture of nucleic acids one or more non-photoreactive placeholding pyrimidines with a photoreactive pyrimidine;

    g) contacting said candidate photoaptamer(s) with said target wherein a candidate photoaptamer-target complex is formed;

    h) irradiating said candidate photoaptamer-target complex;

    i) determining whether said candidate photoaptamer-target complex has photocrosslinked;

    j) repeating (f)-(i) as desired; and

    k) identifying at least one photoaptamer to the target.

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