Application of coding multiple small fragment RNA vector in gene interference and gene activation

A technology of gene interference and small fragments, applied in the field of RNA interference and RNA activation technology, can solve the problems of unsatisfactory effect, not enough to achieve the expected purpose, and limited effect, so as to reduce the cost and cycle of scientific research and reduce the difficulty of screening

Inactive Publication Date: 2008-07-02
WUCHANG SHIPYARD
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Problems solved by technology

[0006] In addition, in many application environments, "RNA interference" or "RNA activation" that only targets a single site or is used alone often has unsatisfactory or limited effects, especially when there are multiple links in certain metabolic pathways, or multiple When there are alternative pathways, the regulation of a single link or just one pathway is often not enough to achieve the desired purpose.

Method used

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  • Application of coding multiple small fragment RNA vector in gene interference and gene activation
  • Application of coding multiple small fragment RNA vector in gene interference and gene activation
  • Application of coding multiple small fragment RNA vector in gene interference and gene activation

Examples

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Embodiment

[0078] A plasmid vector encoding four small fragments of RNA is prepared and applied to simultaneously interfere with Survivin gene and activate E-cadherin gene.

[0079] 1. RNA regulation mechanism and target site design of each gene

[0080] In this example, the plasmid vector encoding four small fragments of RNA is applied to regulate the Surviyin and E-cadherin genes in the prostate cancer cell line PC-3, because resistance to the Surviyin gene in PC-3 will lead to apoptosis and to The expression and activation of E-cadherin gene will inhibit cell proliferation. In this example, RNA interference is applied to Survivin gene, and RNA activation is applied to E-cadherin, so as to control the PC-3 cell carcinoma by inducing apoptosis and resisting its proliferation. cell growth. Each gene is attacked by two target sites that have been published in the literature. shRNA1 and shRNA2 are used for the interference of Survivin, and shRNA3 and shRNA4 are used for the activation of...

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Abstract

The invention provides an application method of a vector capable of simultaneously coding multiple short-hairpin RNAs (shRNAs) for activating the same or different genes, or for simultaneously performing gene interference or gene activation to different genes. The method comprises the following steps of: (1) determining a regulation scheme of a target gene, designing the number of the shRNA corresponding to each gene, and selecting the target site; (2) synthesizing DNA template for coding shRNAs, and constructing a multiple shRNAs expression vector; and (3) applying the constructed multiple shRNAs expression vector to am experimental subject with a proper way as required. The invention provides a convenient and effective application scheme for applying the specific regulation mechanism of shRNA to scientific research and clinics.

Description

technical field [0001] The utility model relates to biotechnology, in particular to RNA interference and RNA activation technology. In particular, it relates to the application of vectors encoding multiple (2 or more) small fragments of RNA for activating the same or different genes, or for respectively performing gene interference and gene activation on different genes. Background technique [0002] RNA interference technology was discovered in 1998, and was successfully used in mammalian cells in May 2001, starting the wide application of RNA interference technology. RNA interference technology uses 21nt or 29nt double-stranded RNA to block the target gene to interfere with the target gene. RNA interference is the oldest genetic immune surveillance mechanism in life. Life maintenance and health, anti-virus invasion, anti-tumor occurrence, and maintenance of genetic stability may all be achieved by RNA interference. Therefore, the research on RNA interference and the appl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/63A61K48/00C12N15/113
Inventor 封江南李超黄俊吴琼杉余铭恩孙永林廖桂红严俊
Owner WUCHANG SHIPYARD
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