US 12,486,530 B2
Multiplexed imaging using strand displacement
Xi Chen, West Newton, MA (US); and Stephanie Hennek, Medford, MA (US)
Assigned to Ultivue, Inc., Cambridge, MA (US)
Filed by Ultivue, Inc., Cambridge, MA (US)
Filed on Aug. 29, 2023, as Appl. No. 18/457,805.
Application 18/457,805 is a division of application No. 17/341,490, filed on Jun. 8, 2021, granted, now 11,773,428.
Application 17/341,490 is a division of application No. 16/117,194, filed on Aug. 30, 2018, granted, now 11,060,132, issued on Jul. 13, 2021.
Application 16/117,194 is a continuation of application No. PCT/US2017/032398, filed on May 12, 2017.
Claims priority of provisional application 62/343,791, filed on May 31, 2016.
Claims priority of provisional application 62/336,676, filed on May 15, 2016.
Prior Publication US 2023/0407371 A1, Dec. 21, 2023
Int. Cl. C12Q 1/6806 (2018.01); A61K 47/68 (2017.01); C07K 16/00 (2006.01); C12Q 1/6855 (2018.01); C12Q 1/6869 (2018.01); G01N 33/58 (2006.01)
CPC C12Q 1/6806 (2013.01) [A61K 47/68 (2017.08); C07K 16/00 (2013.01); C12Q 1/6855 (2013.01); C12Q 1/6869 (2013.01); G01N 33/58 (2013.01); C12Q 2525/191 (2013.01); G01N 2458/10 (2013.01)] 10 Claims
OG exemplary drawing
 
1. A method for imaging of at least one target in a sample comprising:
a) providing at least one target-recognizing antibody bound to an oligonucleotide hybridized to a docking strand but not over the full length, wherein a portion of the docking strand that is not complementary to the oligonucleotide comprises a constant domain and a target-specific domain;
b) providing a blocker strand that hybridizes to the docking strand, wherein the blocker strand comprises a constant domain and a target-specific domain that are complementary to the constant domain and the target-specific domain of the docking strand respectively,
c) providing an imager strand that hybridizes to the docking strand oligonucleotide at least in the region of the target-specific region, wherein the imager strand is labeled with a detectable label and allowing the imager strand to hybridize to the docking strand, displacing the blocker strand;
d) optionally providing a bouncer strand that hybridizes to the constant domain of the blocker domain,
e) optionally, if multiple targets are being imaged, providing blocker strands for all other targets, excluding the blocker strand containing the target-specific domain used in step b;
f) imaging the at least one target;
g) optionally removing the imager strand from the docking strand by providing the blocker strand used in step b; and,
h) optionally repeating steps a-g or any subset thereof if multiple targets are being imaged.