Small regulatory RNAs and methods of use
First Claim
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1. A method of identifying a full length small RNA from a signature sequence RNA molecule comprising:
- a. providing a genomic DNA database;
b. identifying said signature sequence of said small RNA molecule from said database using MPSS method, wherein said signature sequence comprises a portion of a full sequence of said small RNA molecule, wherein said small RNA molecule comprises about 15 to about 30 nucleotides;
c. comparing said signature sequence to said genomic database;
d. identifying one or more genomic regions that indicate identity with said signature sequence; and
e. extending said signature sequence by a necessary number of nucleotides to obtain said full sequence of said small RNA molecule.
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Abstract
The present invention relates to unique small ribonucleic acid molecules, for example siRNAs and miRNAs, identified and isolated using MPSS. Specifically, the invention is directed to the identification of a library of unique small RNA sequences from Arabidopsis thaliana. In another aspect, the small RNA sequences themselves are useful for performing biological functions, such as for example, RNA interference.
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Citations
36 Claims
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1. A method of identifying a full length small RNA from a signature sequence RNA molecule comprising:
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a. providing a genomic DNA database;
b. identifying said signature sequence of said small RNA molecule from said database using MPSS method, wherein said signature sequence comprises a portion of a full sequence of said small RNA molecule, wherein said small RNA molecule comprises about 15 to about 30 nucleotides;
c. comparing said signature sequence to said genomic database;
d. identifying one or more genomic regions that indicate identity with said signature sequence; and
e. extending said signature sequence by a necessary number of nucleotides to obtain said full sequence of said small RNA molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A library of small RNA signature sequences from Arabidopsis thaliana comprising a plurality of sequences selected from the group consisting of SEQ ID NOs:
- 1-185,396.
- View Dependent Claims (9)
- 10. An isolated small RNA molecule comprising a nucleic acid sequence having from about 15 to about 30 nucleotides, wherein the nucleic acid is sufficiently complementary to a plant gene to down-regulate the plant gene by RNA interference.
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29. An expression vector comprising a nucleic acid sequence encoding a nucleic acid having at least 75% homology to a member selected from the group consisting of SEQ ID NO. 1-185,413, wherein the expression vector comprises a transcription initiation region;
- a transcription termination region; and
wherein said nucleic acid sequence is operably linked to said initiation region and said termination region. - View Dependent Claims (30)
- a transcription termination region; and
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31. A method for identifying small RNA molecules that are conserved across more than one species, the method comprising:
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a) creation of a genome-wide small RNA library for a first species;
b) creation of a genomic library or genomic-wide small RNA library for a second species;
c) comparing the library of the first species to the library of the second species; and
d) identifying small RNA molecules found in both the first library and the second library. - View Dependent Claims (32, 33)
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34. A method for identifying small RNA molecules that are unique to a single species, the method comprising:
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a) creation of a genome-wide small RNA library for a first species;
b) creation of a genomic library or genomic-wide small RNA library for a second species;
c) comparing the library of the first species to the library of the second species; and
d) identifying small RNA molecules found in only one of said libraries. - View Dependent Claims (35, 36)
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Specification