Fractional initiator hybridization chain reaction
The present disclosure relates to methods and compositions involving HCR reactions that involve initiators that are split into two or more parts. Effective HCR is dependent upon two or more of these split initiators being brought into proximity (e.g., via binding events mediated by a target) such that a full initiator is formed that is capable of triggering HCR signal amplification.
1. A method, comprising:
providing:
a first hairpin monomer,
a second hairpin monomer,
a first fractional initiator probe comprising a first fractional initiator comprising a sequence capable of binding to the first or second hairpin monomer,
a second fractional initiator probe comprising a second fractional initiator comprising a sequence capable of binding to the first or second hairpin monomer adjacent to the region where the first fractional initiator binds, and
a target molecule;
incubating to allow for binding; and
detecting a signal.
2. The method of claim 1 , wherein the target molecule comprises a nucleotide.
3. The method of claim 1 , wherein the first fractional initiator probe and the second fractional initiator probe are combined with the target molecule.
4. The method of claim 3 , wherein unbound first fractional initiator probe and unbound second fractional initiator probe are removed from a sample containing the target molecule by a first wash.
5. The method of claim 4 , wherein following the first wash, colocalized first and second fractional initiator probes on the target molecule are bound by the first hairpin monomer, which is then bound by the second hairpin monomer, resulting in HCR amplification.
6. The method of claim 5 , wherein unbound hairpin monomer is washed from the sample containing the target molecule.
7. The method of claim 1 , wherein an alternating cascade of the first hairpin monomer binding and the second hairpin monomer binding provides a polymerization event.