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Method for applying miRNA clusters in changing cell fates

A microRNA, cell fate technology, applied to cells modified by the introduction of foreign genetic material, introduction of foreign genetic material using a vector, recombinant DNA technology, etc., can solve problems such as few researches

Inactive Publication Date: 2011-09-28
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little research has been done on this type of cell fate change

Method used

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  • Method for applying miRNA clusters in changing cell fates
  • Method for applying miRNA clusters in changing cell fates
  • Method for applying miRNA clusters in changing cell fates

Examples

Experimental program
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Embodiment 1

[0065] 1. Design of carrier primers for miRNA or miRNA clusters;

[0066] Obtain miRN cluster information from http: / / www.mirbase.org / (eg Figure 1a miRNA clusterA, miRNA cluster B, and miRNA cluster C respectively represent miR-200b-429, miR-106a-363 ​​and miR-302-367), and the sequence of the miR-200b-429 clone region is SEQ ID NO: 1, miR- The sequence of the cloning region of 106a-363 ​​is SEQ ID NO: 2, and the sequence of the cloning region of miR-302-367 is SEQ ID NO: 3. By designing specific primers to amplify miRNA or miRNA clusters, the genome includes about 200 upstream and downstream sequence of nucleotides.

[0067] The nucleotide sequence of the upstream primer of miR-200b-429 is shown in SEQ ID NO: 4;

[0068] The nucleotide sequence of the downstream primer of miR-200b-429 is shown in SEQ ID NO: 5;

[0069] The nucleotide sequence of the upstream primer of miR-106a-363 ​​is shown in SEQ ID NO: 6;

[0070] The nucleotide sequence of the downstream primer of mi...

Embodiment 2

[0087] Cluster C miRNA cluster can improve the induction efficiency of human cells, generally, the induction efficiency of human pluripotent cells is lower. Under the condition of not adding the oncogene c-Myc, the efficiency is only about 1 / 10,000 to 1 / 100,000. The same experimental method as in Example 1, except that the mouse genome and mouse embryonic fibroblasts were replaced with human genome and human fibroblasts. Depend on Figure 10 The data shows that Cluster C miRNA clusters can significantly promote the change of human cell fate. This technique is conserved in different species, and the ClusterCmiRNA cluster can also improve the induction efficiency of human cells.

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Abstract

The objective of the invention is to provide a method for applying miRNAs or miRNA clusters in changing cell fates by promoting the formation of induced pluripotent stem cells. The method comprises a first step of cloning genes of miRNAs or miRNA clusters in a donor genome and obtaining a donorcell, a second step of changing the cell fate of the donorcell in step 1 by using the miRNAs or miRNA clusters in step 1, and a third step of identifying the cell obtained in step 2. Compared to the prior art, stem cells are induced by miRNAs or miRNA clusters in the invention instead of by oncogenes c-Myc, thereby reducing potential carcinogenicity, accelerating the change of cell fates and enabling a high induction efficiency and high-quality induced pluripotent stem cells with ability of germ line transmission to be obtained.

Description

technical field [0001] The present invention relates to a method of using microribonucleic acid clusters for changing the fate of cells. Background technique [0002] my country is a country with a large population in the world. Every year, organ defects, failures, and dysfunctions caused by trauma, disease, aging, and genetics also rank first in the world. Therefore, the research on stem cells and regenerative medicine has attracted the general attention of quite a few scientific research institutes and all walks of life. [0003] Cell transplantation therapy is an important direction of regenerative medicine research. Cell transplantation of specific types of cells may be used to treat heart damage, nervous system degenerative diseases, spinal cord injuries, kidney failure, blood system diseases and so on. However, cell transplantation therapy faces many difficult problems such as allogeneic rejection and limited cell sources. [0004] Stem cells are a kind of pluripotent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N5/10
Inventor 裴端卿米格尔·安格尔·艾斯特班鲍习琛廖宝剑张必良阿塔那希奥斯·泽沃伊丽斯
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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