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Microrna and uses thereof

A sequence and nucleic acid technology, applied in the field of microRNA and its application, can solve problems such as difficult identification of miRNAs

Inactive Publication Date: 2007-09-05
ROSETTA GENOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to their small size, miRNAs have been difficult to identify using standard methodologies

Method used

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  • Microrna and uses thereof
  • Microrna and uses thereof
  • Microrna and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0142] miRNA prediction

[0143] We used two in silico methods similar to those used to predict miRNAs described in U.S. Patent Application Nos. 60 / 522,459, 10 / 709,577, and 10 / 709,572, the contents of which are incorporated herein by reference, for potential miRNA-encoding gene pairs. The entire human genome was examined. Briefly, nonprotein-coding regions of the entire human genome were scanned for hairpin structures. Predicted hairpin structures and potential miRNAs were scored according to thermodynamic stability as well as structural and contextual features. Validated miRNA calibration algorithms from the Sanger database were used.

[0144] 1. First Screening

[0145] Tables 11A-11C show the sequence ("precursor sequence"), sequence identifier ("PRECUR SEQ-ID"), and organism of origin ("GAM organism") and Predicted miRNAs ("GAM names"). Table 12 shows the sequence ("GAM RNA sequence") and sequence identifier ("GAM SEQ-ID") of each miRNA ("GAM na...

Embodiment 2

[0154] Prediction of target genes

[0155] The 2 in silico screens from Example 1 were then combined using in silico methods similar to those described in U.S. Patent Application Nos. 60 / 522,459, 10 / 709,577, and 10 / 709,572 (the contents of which are incorporated herein by reference) for predicting The predicted miRNAs are used to predict target genes and their binding sites.

[0156] 1. The first screening

[0157] Tables 13A-13C list the predicted target gene (“Target”) and binding site sequence (“Target Binding Site Sequence”) and binding site sequence identifier (“Target Binding Site SEQ-ID"), and the organism of origin of the target ("target organism"). Table 14 lists diseases ("Disease Names") associated with target genes ("Disease-Associated Target Genes"). Table 15 lists miRNA sequence identifiers ("SEQ ID NOs of GAMS associated with diseases") and diseases associated with miRNAs based on target genes ("Disease Name"). The sequences of the bind...

Embodiment 3

[0164] Validation of miRNA

[0165] 1. Expression Analysis - Group 1

[0166] To determine predicted hairpin structures and miRNAs on 0, we used high-throughput microarrays similar to those described in US Patent Application Nos. 60 / 522,459, 10 / 709,577 and 10 / 709,572, the contents of which are incorporated herein by reference Arrays detect expression in various tissues. For each predicted precursor miRNA, mature miRNAs derived from the two stems of the hairpin were examined.

[0167] Table 2 shows the hairpin structure ("HID") of the second predicted panel validated by detecting the expression of the associated miRNA ("MID"), and the code of the tissue ("tissue") in which expression was detected. The codes for the tissues and diseases of Table 2 are listed in Table 7. Some of the tissues tested are cell lines. The lung cancer cell line (H1299) with / without P53:H1299 has mutated P53. Cell lines were transfected with such a construct with P53, whic...

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Abstract

Described herein are novel polynucleotides associated with prostate and lung cancer. The polynucleotides are miRNAs and miRNA precursors. Related methods and compositions that can be used for diagnosis, prognosis, and treatment of those medical conditions are disclosed. Also described herein are methods that can be used to identify modulators of prostate and lung cancer.

Description

Field of Invention [0001] The present invention generally relates to microRNA molecules and various nucleic acid molecules associated therewith or derived therefrom. Background of the Invention [0002] MicroRNAs (miRNAs) are short RNA oligonucleotides of approximately 22 nucleotides involved in gene regulation. MicroRNAs regulate gene expression by targeting mRNA for cleavage or translational repression. Although miRNAs are present in a wide range of species, including C. elegans, Drosophila, and humans, they have only recently been identified. More importantly, the role of miRNAs in disease development and progression has only recently begun to be understood. [0003] Due to their small size, miRNAs have historically been difficult to identify using standard methodologies. A limited number of miRNAs have been identified by extracting large amounts of RNA. miRNAs that contribute to the display of visually discernible phenotypes have also been ident...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor I·本特维奇A·阿夫尼尔Y·卡洛夫R·阿哈罗诺夫
Owner ROSETTA GENOMICS
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