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MiR-16 regulated genes and pathways as targets for therapeutic intervention

A gene and pathway technology, applied in biochemical equipment and methods, microbial assay/inspection, DNA/RNA fragments, etc., can solve problems such as unknown details of regulatory pathways and networks

Inactive Publication Date: 2010-01-20
ASURAGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current limitation of this approach is that, as noted above, the details of the regulatory pathways and networks affected by any given miRNA remain largely unknown

Method used

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  • MiR-16 regulated genes and pathways as targets for therapeutic intervention
  • MiR-16 regulated genes and pathways as targets for therapeutic intervention
  • MiR-16 regulated genes and pathways as targets for therapeutic intervention

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Experimental program
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Effect test

preparation example Construction

[0235] B. Preparation of nucleic acid

[0236] Nucleic acids can be prepared by any technique known to those of ordinary skill in the art, such as chemical synthesis, enzymatic production or biological production. It is specifically considered that the miRNA probe of the present invention is chemically synthesized.

[0237] In some embodiments of the invention, miRNA is recovered or isolated from a biological sample. The miRNA can be recombinant, or it can be the cell's natural or endogenous miRNA (produced from the cell's genome). It is considered that biological samples can be processed to enhance the recovery of small RNA molecules such as miRNA. This method is described in US Patent Application Serial No. 10 / 667,126, which is specifically incorporated herein by reference. Generally, these methods involve lysing cells with a solution with guanidine salt and detergent.

[0238] Alternatively, nucleic acid synthesis can be performed according to standard methods. See, for examp...

Embodiment 1

[0279] Analysis of gene expression after HSA-MIR-16 transfection

[0280] miRNAs are thought to affect gene expression mainly at the translation level. Translation regulation leads to an upward or downward change in protein expression, which can lead to changes in the activity and expression of downstream gene products and genes that are regulated by these proteins. These regulatory effects will appear as changes in the global mRNA expression profile. In addition, it has recently been reported that, in some cases, miRNAs can reduce the mRNA levels of their direct targets (Bagga et al., 2005; Lim et al., 2005), and this change can be observed in microarray gene expression analysis. Microarray gene expression analysis was performed to identify genes that are misregulated by hsa-miR-16.

[0281] For each of the three time points, the synthetic precursor miR-16 (Ambion) was reverse stained into quadruplicate samples of A549 cells. Cells were transfected with siPORT NeoFX (Ambion) ac...

Embodiment 2

[0290] Cellular pathways affected by HSA-MIR-16

[0291] The misregulation of gene expression by hsa-miR-16 (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-16 expression. Use Ingenuity Pathways Analysis ( Systems, Redwood City, CA) performs cell path analysis. The most affected pathways after hsa-miR-16 overexpression in A549 cells are shown in Table 2.

[0292] Table 2. Functional cellular pathways that are significantly affected after hsa-miR-16 is overexpressed in human cancer cells

[0293]

[0294] These data prove that hsa-miR-16 directly or indirectly affects the expression of numerous metabolism, cell proliferation, cell development and cell cycle related genes, and therefore mainly affects the functional pathways related to cell growth, developme...

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Abstract

The present invention concerns methods and compositions for identifying genes or genetic pathways modulated by miR-16, using miR-16 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

[0001] This application claims priority for US provisional application number 60 / 882,758 filed on December 29, 2006 and PCT application PCT / US07 / 87038 filed on December 10, 2007, both of which are incorporated herein by reference in their entirety. [0002] This application relates to US patent application serial number 11 / 141,707 filed on May 31, 2005 and serial number 11 / 273,640 filed on November 14, 2005, each of which is incorporated herein by reference in its entirety. Technical field [0003] The invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to methods and compositions for treating diseases or conditions that are affected by miR-16 microRNA, microRNA expression, and genes and cell pathways directly and indirectly regulated by them. Background technique [0004] In 2001, several organizations used cloning methods to isolate and identify a large group of "microRNA" (miRNA) from C.elegans, Drosophila and humans...

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 麦克·拜罗姆卢娜·帕特拉瓦拉查尔斯·D·约翰逊大卫·布朗安德里斯·G·巴德
Owner ASURAGEN
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