Methods and materials using signaling probes
First Claim
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.
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Accused Products
Abstract
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.
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Citations
18 Claims
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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:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method of isolating cells each expressing RNAs from two or more RNA expression libraries, comprising the steps of:
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introducing into cells a plurality of DNAs encoding a first RNA expression library, wherein each DNA further encodes a first nucleic acid tag sequence, and wherein at least a subset of the plurality of DNAs encodes the same first nucleic acid tag sequence; introducing into the cells a plurality of DNAs encoding a second RNA expression library, wherein each DNA further encodes a second nucleic acid tag sequence, and wherein at least a subset of the plurality of DNAs encodes the same second nucleic acid tag sequence; exposing the cells to a first signaling probe that produces a detectable signal upon hybridization to said first nucleic acid tag sequence; exposing the cells to a second signaling probe that produces a detectable signal upon hybridization to said second 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 for each of the first and second signaling probes.
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Specification