Abstract: The present disclosure relates to an HCR lateral flow device for amplified detection of a target in a sample and methods of using the same. In some embodiments, the device and methods of use combine the sensitivity of HCR signal amplification with the simplicity of the lateral flow assay format to facilitate user-friendly amplified detection of a target in a sample.

Abstract: Programmable guide RNAs (gRNAs) play a central role in the CRISPR revolution sweeping biology and medicine by directing the function of a Cas protein effector to a target gene of choice. To achieve programmable control over regulatory scope, the activity of a conditional guide RNA (cgRNA) depends on the presence or absence of an RNA trigger, allowing for cell-selective regulation of CRISPR/Cas function. Unlike a standard gRNA, a cgRNA is programmable at multiple levels, with the target-binding sequence controlling the target of Cas activity (edit, silence, induce, or bind a gene of choice) and the trigger binding sequence controlling the scope of Cas activity. cgRNA mechanisms that are allosteric allow for independent design of the target and trigger sequences, providing the flexibility to select the regulatory target and scope independently.

Abstract: Methods of analysis of a sample using hybridization chain reaction (HCR) are provided herein. Some embodiments involve one, two, or all three of the following aspects: 1) repeated signal detection, 2) overlapping binding sites, and 3) catalytic reporter deposition (CARD). Compositions and kits relating to these are also provided. Some embodiments encompass a method for repeated signal detection with reporter-labeled HCR hairpins involving providing a sample possibly containing one or more targets as well as possibly other molecules that are not targets, providing one or more probe sets each comprising either: a) one or more HCR initiator-labeled probes, or b) one or more probe units each comprising two or more HCR fractional initiator probes, providing one or more HCR amplifiers (each labeled with one or more reporters), detecting one or more signals from one or more reporters.

Abstract: Programmable guide RNAs (gRNAs) play a central role in the CRISPR revolution sweeping biology and medicine by directing the function of a Cas protein effector to a target gene of choice. To achieve programmable control over regulatory scope, the activity of a conditional guide RNA (cgRNA) depends on the presence or absence of an RNA trigger, allowing for cell-selective regulation of CRISPR/Cas function. Unlike a standard gRNA, a cgRNA is programmable at multiple levels, with the target-binding sequence controlling the target of Cas activity (edit, silence, induce, or bind a gene of choice) and the trigger binding sequence controlling the scope of Cas activity. cgRNA mechanisms that are allosteric allow for independent design of the target and trigger sequences, providing the flexibility to select the regulatory target and scope independently.

Abstract: Methods of analysis of a sample using hybridization chain reaction (HCR) are provided herein. Some embodiments involve one, two, or all three of the following aspects: 1) repeated signal detection, 2) overlapping binding sites, and 3) catalytic reporter deposition (CARD). Compositions and kits relating to these are also provided. Some embodiments encompass a method for repeated signal detection with reporter-labeled HCR hairpins involving providing a sample possibly containing one or more targets as well as possibly other molecules that are not targets, providing one or more probe sets each comprising either: a)one or more HCR initiator-labeled probes, or b) one or more probe units each comprising two or more HCR fractional initiator probes, providing one or more HCR amplifiers (each labeled with one or more reporters), detecting one or more signals from one or more reporters.

Abstract: Nucleic acids (DNA and RNA) provide a versatile platform for engineering synthetic biology in a variety of technology areas including medicine, science, agriculture, and energy. In many settings, degradation of nucleic acid molecules poses a significant engineering challenge as the molecules do not function if they have been degraded. In some embodiments, nucleic acid protective elements (PELs) are used to protect chemically synthesized or expressed nucleic acid molecules from degradation. PELs may be derived from all or part of a viral xrRNA sequence and/or structural motif, PELs may include rationally designed sequences and/or structural motifs, PELs may be engineered using directed evolution, and in some embodiments, PELs comprise a mixture of biologically derived, rationally designed sequence and/or structural motifs, and/or sequences and/or structural motifs that are engineered by directed evolution.

Abstract: 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.

Abstract: 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.

Abstract: The present disclosure relates to methods involving HCR reactions that involve using trigger oligos to activate probes that initiate HCR.

Abstract: 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.

Abstract: 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.

Abstract: Described herein are conditional guide RNAs that change their activity status depending on the presence or absence of an input target, forming a complex with an RNA-guided effector and conditionally performing a downstream function on a target nucleic acid. Methods for conditionally performing a downstream function on the target nucleic acid using conditional guide RNAs are also described.

Abstract: The present disclosure relates to methods involving HCR reactions that involve using trigger oligos to activate probes that initiate HCR.

Abstract: 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.

Abstract: The present subject matter relates to the use conditional hairpins, such as, but not limited to shRNAs. The conditional formation of these structures can allow for further events, such as gene silencing (in some embodiments).

Abstract: The present invention relates generally to programming of biomolecular self-assembly pathways and related methods and constructs for self-assembly of prescribed two and three-dimensional structures.

Abstract: The present invention relates generally to programming of biomolecular self-assembly pathways and related methods and constructs. A versatile nucleic acid hairpin motif for programming biomolecular self-assembly pathways for a wide variety of dynamic functions, reaction graphs for specifying pathways, and methods of using the hairpin motif are provided.

Abstract: Methods and systems for designing the sequences of multiple nucleic acid strands intended to hybridize in solution via a prescribed reaction pathway are described. Sequence design is formulated as a multistate optimization problem using a set of target test tubes containing different subsets of the strands to represent reactant, intermediate, and product states of the system. Each target test tube contains a set of desired “on-target” complexes, each with a target secondary structure and target concentration, and a set of undesired “off-target” complexes, each with vanishing target concentration. Optimization of the equilibrium ensemble properties of the target test tubes may implement both a positive design paradigm, explicitly designing for on-pathway states, and a negative design paradigm, explicitly designing against off-pathway states.

Abstract: The present embodiments relate to engineering imaging probes based on “triggered molecular geometry.” Upon detection of a molecular signal, nucleic acid hairpin monomers assemble an imageable molecular shape with prescribed geometry. In some embodiments the prescribed shape can be imaged directly. In some embodiments, the prescribed shape can serve as a spatial organizer or amplification scheme for other imaging entities, such as fluorophore and fluorescent proteins.

Abstract: The present application relates to the use of hybridization chain reaction (HCR) to form double stranded RNA polymers in the presence of a target, such as a nucleic acid associated with a disease or disorder. The RNA polymers are preferably able to activate the RNA-dependent kinase PKR. Activation of PKR via RNA-HCR can be used to treat a wide variety of diseases and disorders by specifically targeting diseased cells.