Caged RNAs and methods of use thereof

Abstract

Methods of using labeled interfering RNAs to detect and / or quantitate target mRNAs in cells are provided. Related compositions, systems, and kits are also provided. Caged interfering RNAs (e.g., photoactivatable interfering RNAs), methods of using such caged RNAs, and related systems and kits are also provided. Caged RNAs capable of repressing translation of a target mRNA or silencing transcription of a target gene are also provided, along with related methods, systems, and kits. Methods and compositions for introducing RNAs into cells, using RNAs covalently associated with protein transduction domains and / or lipids, are provided. Also provided are methods and compositions for selectively attenuating expression of a target mRNA by controlling expression of an interfering RNA, an RNA capable of initiating translation repression, or an RNA capable of initiating transcriptional silencing.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to and benefit of U.S. Provisional Patent Application No. 60 / 572,564, filed May 18, 2004, entitled “RNAi-based Sensors, Caged RNAs, and Methods of Use Thereof” by Nguyen et al., and is a continuation-in-part of U.S. patent application Ser. No. 10 / 716,393, filed Nov. 17, 2003, entitled “RNAi-based Sensors, Caged Interfering RNAs, and Methods of Use Thereof” by Nguyen and McMaster, which claims priority to and benefit of the following prior provisional patent applications: U.S. Ser. No. 60 / 427,664, filed Nov. 18, 2002, entitled “Photo Activated Sensors, Regulators and Compounds” by Nguyen and McMaster, U.S. Ser. No. 60 / 436,855, filed Dec. 26, 2002, entitled “Caged Sensors, Regulators and Compounds and Uses Thereof” by Nguyen and McMaster, U.S. Ser. No. 60 / 439,917, filed Jan. 13, 2003, entitled “Caged Sensors, Regulators and Compounds and Uses Thereof” by Nguyen and McMaster, U.S. Ser. No. 60 / 451,177, filed...

AI Technical Summary

Problems solved by technology

Furthermore, current methods typically involve multi-step processing that contributes to high intra- and inter-assay variation.

Consequently, current methods do not provide live, dynamic, and location-specific imaging and measurements of mRNA, because the cells are exposed to environmental changes.

However, the complexity and richness in cellular assay data sets, compared to genomics and proteomics studies, will provide scientists with unparalleled tools to aid discovery efforts throughout the discovery process and for basic research applications.

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