Multiplexed single molecule RNA fish using DNA nanostructures

EP4771177A1Pending Publication Date: 2026-07-08LMU LUDWIG-MAXIMILIANS-UNIVERSITÄT
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Patent Information

Application Number
EP2024782293
Authority / Receiving Office
EP · EP
Patent Type
Applications
Current Assignee / Owner
Priority Date
2023-09-28
Filing Date
2024-09-27
Publication Date
2026-07-08

AI Technical Summary

Technical Problem

Current RNA fluorescence in situ hybridization (FISH) techniques are limited in their ability to simultaneously detect multiple RNA species with spatial context, due to the constraints of fluorescence microscopy and the need for sequential hybridization steps.

Method used

The development of multiplexed single molecule RNA FISH using DNA nanostructures, which involves probes comprising a marker molecule, a targeting molecule with a spacer sequence, and a target binding sequence, allowing for simultaneous detection of multiple RNAs with enhanced signal strength and reduced non-specific interactions.

Benefits of technology

This approach enables the simultaneous detection of up to 48 RNA species with high specificity and sensitivity, retaining spatial context and reducing the complexity of experimental procedures, making it suitable for non-specialist laboratories and clinical settings.

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Abstract

The present invention relates to a probe for RNA fluorescence in situ hybridization (FISH) comprising: a marker molecule, a targeting molecule comprising 22 to 55 nt comprising a spacer sequence, and a target binding sequence capable of binding to a target molecule, wherein the marker molecule is connected to the targeting molecule via the spacer sequence The present invention also relates to a composition for RNA FISH comprising a set of probes for RNA FISH, wherein the set of probes comprises at least 1 probe for RNA FISH, preferably at least 5 probes for RNA FISH, more preferably at least 10 probes for RNA FISH, wherein the set of probes comprises at least one probe. The invention further relates to a method for designing a probe for RNA FISH for analyzing a sample, wherein the method comprises providing a probe for RNA FISH, wherein the method comprises detecting at least one target molecule in situ in the sample using the probe. The present invention further relates to a kit for use in a molecular analysis, the comprising a probe according to any of the preceding probe claims, or a composition according to any of the preceding composition claims, and at least one buffer for hybridization comprising at least one of: fixation buffer component, wash buffer, and hybridization buffer.
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