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miR-20 regulated genes and pathways as targets for therapeutic intervention

A mir-21 and pathway technology, applied in the fields of molecular biology and medicine, can solve problems such as limitations

Inactive Publication Date: 2010-01-06
ASURAGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, this represents an important limitation to cancer therapy in which miR-21 may play a role

Method used

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  • miR-20 regulated genes and pathways as targets for therapeutic intervention
  • miR-20 regulated genes and pathways as targets for therapeutic intervention
  • miR-20 regulated genes and pathways as targets for therapeutic intervention

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0310] Example 1: Analysis of gene expression after HSA-MIR-21 transfection

[0311] It is believed that miRNAs regulate gene expression by binding to target mRNA transcripts and (1) initiating transcript degradation or (2) altering the translation of proteins from the transcript. Translational regulation that leads to up- or down-regulation of protein expression can produce changes in downstream gene products and gene activity and expression, and translation regulation is in turn regulated by these proteins. These numerous regulatory effects are shown as changes in the overall mRNA expression profile. A microarray gene expression analysis was performed to identify genes misregulated by hsa-miR-21 expression.

[0312] Synthetic precursor-miR-21 (Ambion) or two negative control miRNAs (precursor-miR-NC1, Ambion catalog number AM17110 and precursor-miR-NC2, Ambion, catalog number AM17111) reverse infection into quadruplicate In the A549 cell sample, each is three time points. Cell...

Embodiment 2

[0314] Example 2: Cellular pathways affected by HSA-miR-21

[0315] The misregulation of gene expression by hsa-miR-21 (Table 1) affects many cellular pathways that represent potential therapeutic targets for the control of cancer and other diseases and disorders. The inventors determined the identity and nature of the cytogenetic pathways affected by the regulatory cascade induced by hsa-miR-21 expression. Cell pathway analysis uses Ingenuity Pathways Analysis (Version 4.0, Ingenuity Systems, Redwood City, CA). The change of a specific pathway was determined by Fisher's exact test (Fisher, 1922). The pathways most significantly affected by hsa-miR-21 overexpression in A549 cells are shown in Table 2.

[0316] These data prove that hsa-miR-21 directly or indirectly affects the expression of many cancers, cell proliferation, cell development, cell signal transduction and cell cycle-related genes, and therefore mainly affects functions related to cell growth, cell development and ...

Embodiment 3

[0317] Example 3: Predicted HSA-MIR-21 gene target

[0318] The gene target that binds hsa-miR-21 and is regulated by hsa-miR-21 uses a proprietary algorithm miRNATarget TM (Asuragen) predicts that this algorithm is an implementation of the method suggested by Krek et al. (2005). The predicted target genes are shown in Table 3. The predicted gene targets showing changes in the mRNA expression level in human cancer cells transfected with the precursor-miR hsa-miR-21 are shown in Table 4.

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Abstract

The present invention concerns methods and compositions for identifying genes or genetic pathways modulated by miR-21, using miR-21 to modulate a gene or gene pathway, using this profile in assessing the condition of a patient and / or treating the patient with an appropriate miRNA.

Description

Background of the invention [0001] I. Technical Field [0002] The invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to methods for treating diseases or conditions that are affected by miR-21 small RNA, small RNA expression, and genes and cellular pathways directly and indirectly regulated by this. [0003] II. Background Technology [0004] In 2001, several groups used cloning methods to isolate and identify a large group of "small RNAs" (miRNAs) from C. elegans, Drosophila and humans (Lagos-Quintana et al., 2001; Lau et al., 2001; Lee and Ambrmbros, 2001). Hundreds of miRNAs have been identified in plants and animals including humans, and they do not seem to have endogenous siRNA. Therefore, although miRNA is similar to siRNA, miRNA is different. [0005] The miRNAs observed so far are approximately 21-22 nucleotides in length, and they are derived from longer precursors transcribed from non-protein-coding genes. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11
CPCC12Q2600/178C12Q2600/112C12Q1/6886C12Q1/6809C12Q2600/118C12Q2525/207
Inventor 安德里斯·G·巴德麦克·拜罗姆查尔斯·D·约翰逊大卫·布朗
Owner ASURAGEN
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