METHODS AND MATERIALS USING SIGNALING PROBES

摘要

The present invention relates to methods of isolating cells, generating cell lines, and detecting RNAs in cells using signaling probes that produce a signal upon hybridization to a target sequence. Other methods that utilize the signaling probe include methods of detecting or quantifying the effect of an agent on RNAs in a cell, methods of quantifying the level of RNA expression, methods for identifying genetic recombinational events in living cells and methods of generating a transgenic animal using the isolated cells. The invention also provides protease probes. Signaling probes and protease probes that form stem-loop structures, three-arm junction structures, and dumbbell structures are provided.

主权项

1. A method of detecting the effect of an agent on the level of an RNA of interest, comprising the steps of:
a) exposing cells capable of expressing said RNA of interest to the agent; b) exposing the cells to a signaling probe that produces a detectable signal upon hybridization with the RNA of interest, wherein said signaling probe comprises a region of nucleotides complementary to the RNA of interest and regions of mutually complementary nucleotides; and c) comparing the signal produced after exposure to the agent to the signal produced in the absence of the agent, to detect the effect of the agent on the level of the RNA of interest.