Provided herein is a cell-free biosensor lateral flow device kit for detection of analytes of interest and methods of using thereof. The device comprises a substrate and a first end, wherein the first end comprises a sample loading portion. The device additionally may comprise a sensor module, wherein the sensor module comprises an allosteric transcription factor regulated in vitro transcription reaction, and a transduction module, wherein the transduction module comprises Bait and Prey nucleic acids which sense the output of the sensor element and a reporter conjugate which accumulates at a test zone when an analyte the sensor element senses is present in the sample.

Provided herein is a cell-free biosensor lateral flow device kit for detection of analytes of interest and methods of using thereof. The device comprises a substrate and a first end, wherein the first end comprises a sample loading portion. The device additionally may comprise a sensor module, wherein the sensor module comprises an allosteric transcription factor regulated in vitro transcription reaction, and a transduction module, wherein the transduction module comprises Bait and Prey nucleic acids which sense the output of the sensor element and a reporter conjugate which accumulates at a test zone when an analyte the sensor element senses is present in the sample.

Disclosed are compositions, systems, kits, and methods for detecting an analyte or target molecule in a sample by regulated in vitro transcription. The compositions, systems, kits, and methods typically comprise and/or utilize one or more components selected from: (a) an RNA polymerase; (b) an allosteric transcription factor (ATT), wherein the ATT binds an analyte or target molecule as a ligand; (c) an engineered transcription template; and/or any combination thereof. The engineered transcription template typically comprises a promoter sequence for the RNA polymerase and an operator sequence for the ATT. The promoter sequence and operator sequence are operably linked to a sequence encoding an RNA, wherein the ATT modulates transcription of the encoded RNA when the ATT binds the analyte or target molecule as a ligand, wherein the transcribed RNA generates a detectable signal in conjunction with a reporter molecule.

Disclosed are compositions, systems, kits, and methods for detecting an analyte or target molecule in a sample by regulated in vitro transcription. The compositions, systems, kits, and methods typically comprise and/or utilize one or more components selected from: (a) an RNA polymerase; (b) an allosteric transcription factor (ATT), wherein the ATT binds an analyte or target molecule as a ligand; (c) an engineered transcription template; and/or any combination thereof. The engineered transcription template typically comprises a promoter sequence for the RNA polymerase and an operator sequence for the ATT. The promoter sequence and operator sequence are operably linked to a sequence encoding an RNA, wherein the ATT modulates transcription of the encoded RNA when the ATT binds the analyte or target molecule as a ligand, wherein the transcribed RNA generates a detectable signal in conjunction with a reporter molecule.

The present invention is directed to a method for immobilizing nucleic molecule on solid support and to a use of a nucleic acid non-immobilized primer in combination with a nucleic acid primer linked to a solid support in said a method.

The present invention is directed to a method for immobilizing nucleic molecule on solid support and to a use of a nucleic acid non-immobilized primer in combination with a nucleic acid primer linked to a solid support in said a method.

Described herein are nucleic acid detection compositions and systems comprising isothermal loop mediated signal amplification (LAMP) and methods of using these LAMP-based compositions and systems.

Described herein are nucleic acid detection compositions and systems comprising isothermal loop mediated signal amplification (LAMP) and methods of using these LAMP-based compositions and systems.

Embodiments of the instant disclosure relate to novel compositions and methods for analyte detection by quantifying transcription of RNA using CRISPR-based approaches. In certain embodiments, constructs of use in assays disclosed herein can include a novel double-stranded DNA sequence having at least one transcriptional promoter, at least one regulatory element, and a target sequence. In some embodiments, constructs disclosed herein can be used in a system for quantifying transcriptional output in a sample in order to detect presence of an analyte. In other embodiments, constructs and systems disclosed herein can be used to measure enzyme activity, to detect and/or quantify at least one agent that modulates transcription, detect and/or quantify analytes (e.g. a contaminant, biomarkers or other agent) or a combination thereof in accordance with assessing health of a subject or assessing environmental conditions or contaminants.

Embodiments of the instant disclosure relate to novel compositions and methods for analyte detection by quantifying transcription of RNA using CRISPR-based approaches. In certain embodiments, constructs of use in assays disclosed herein can include a novel double-stranded DNA sequence having at least one transcriptional promoter, at least one regulatory element, and a target sequence. In some embodiments, constructs disclosed herein can be used in a system for quantifying transcriptional output in a sample in order to detect presence of an analyte. In other embodiments, constructs and systems disclosed herein can be used to measure enzyme activity, to detect and/or quantify at least one agent that modulates transcription, detect and/or quantify analytes (e.g. a contaminant, biomarkers or other agent) or a combination thereof in accordance with assessing health of a subject or assessing environmental conditions or contaminants.