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

A pathway, gene technology, applied in the direction of biochemical equipment and methods, microbial assay/inspection, DNA/RNA fragments, etc., which 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 on cancer therapy in which miR-20 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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0383] Example 1: Analysis of gene expression after HSA-MIR-20a transfection

[0384] 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-20a expression.

[0385] Synthetic Pre-miR-20a (Ambion) was counter-infected into quadruplicate A549 cell samples, each for three time points. Cells were transfected with siPORT NeoFX (Ambion) according to the manufacturer's recommendations with the following parameters: 200,000 cells per...

Embodiment 2

[0388] Example 2: Cellular pathways affected by HSA-MiR-20a

[0389] The misregulation of gene expression by hsa-miR-20a (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-20a expression. Cell pathway analysis uses Ingenuity Pathways Analysis ( Systems, Redwood City, CA). The pathways most significantly affected by hsa-miR-20a overexpression in A549 cells are shown in Table 2 above.

[0390] These data prove that hsa-miR-20a directly or indirectly affects the expression of numerous cell growth, cell proliferation, cell signal transduction and cell development-related genes, and therefore mainly affects the functional pathways related to cell growth, cell development and cell proliferation. These cellular processes all have an integral role in the developmen...

Embodiment 3

[0391] Example 3: Predicted HSA-MIR-20a gene target

[0392] The gene target that binds to hsa-miR-20a and is regulated by hsa-miR-20a uses a proprietary algorithm miRNATarget TM (Asuragen) predicted and shown in Table 3 above.

[0393] The predicted gene targets showing changes in mRNA expression levels in human cancer cells transfected with pre-miR hsa-miR-20a are shown in Table 4 above.

[0394] The predicted gene target of hsa-miR-20a whose mRNA expression level is affected by hsa-miR-20a represents a particularly useful candidate for cancer treatment and other disease treatment by controlling their expression level.

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Abstract

The present invention concerns methods and compositions for identifying genes or genetic pathways modulated by miR-20a, using miR-20a 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-20 microRNA, microRNA expression, and genes and cellular pathways directly and indirectly regulated by them. [0003] II. Background Technology [0004] In 2001, several groups used cloning methods to isolate and identify a large group of "microRNA" (miRNA) 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. See re...

Claims

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Application Information

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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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