Methods and materials using signaling probes

摘要

Methods of isolating cells or generating cell lines comprising the step of exposing the cells to signaling probes that produce a signal upon hybridization to a target sequence, as well as methods of quantifying the level of expression of an RNA of interest, methods for identifying genetic recombinational events in living cells and methods of generating a transgenic animal using the isolated cells. Methods for isolating a plurality of cells encoding a plurality of different RNAs associated with a same nucleic acid tag sequence, comprising the step of exposing the cells to a same signaling probe that produces a detectable signal upon hybridization to the same nucleic acid tag sequence, are also provided. Signaling probes and protease probes that form stem-loop structures, three-arm junction structures, and dumbbell structures may be used in the above methods.

主权项

1. A method of isolating a plurality of cells, wherein a subset of the cells expresses an RNA that is not expressed by another subset of the cells, comprising the steps of:
introducing into cells a plurality of DNAs encoding a plurality of different RNAs, wherein each DNA further encodes a nucleic acid tag sequence, and wherein at least a subset of the plurality of DNAs encodes the same nucleic acid tag sequence; exposing the cells to a signaling probe that produces a detectable signal upon hybridization to said same nucleic acid tag sequence; and isolating, in a single application, the cells that exhibit at least one selected level of fluorescence at a selected wavelength.