MULTIPLEX LABELING OF MOLECULES BY SEQUENTIAL HYBRIDIZATION BARCODING USING PROBES WITH CLEAVABLE LINKERS

摘要

The present invention, among other things, provides technologies for detecting and/or quantifying nucleic acids in cells, tissues, organs or organisms. Through sequential barcoding, the present invention provides methods for high-throughput profiling of a large number of targets, such as transcripts and/or DNA loci. In some embodiments, nucleic acid probes include a signal moiety connected with a binding sequence via a cleavable linker.

主权项

1. A composition, comprising:
a plurality of primary probes, wherein each primary probe comprises:
a primary binding sequence that binds to a complementary target sequence in a target nucleic acid molecule, anda first overhang sequence connected to one end of the primary binding sequence; a first plurality of bridge probes, wherein each bridge probe comprises:
a binding sequence that specifically binds to all or a part of the first overhang sequence of a primary probe of the plurality of primary probes, andone or more readout binding targets connected in series and linked to the binding sequence; a first plurality of readout probes, wherein each readout probe comprises:
a readout binding sequence that specifically binds to a first readout binding target of the one or more readout binding targets of a bridge probe of the first plurality of bridge probes, anda signal moiety linked to the readout binding sequence via a cleavable linker, wherein the signal moiety is capable of emitting a first detectable visual signal upon binding of each readout probe from the first plurality of readout probes to the first readout binding target of one of the one or more readout binding targets.