Method for promoting RNA degradation
A DNA sequence and binding domain technology, applied in the field of genetic engineering, can solve the problem that the degradation effect cannot reach the high-efficiency target, and achieve the effect of improving the ability to degrade RNA and improve the ability of RNA
Inactive Publication Date: 2018-07-24
李燕强
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Problems solved by technology
The degradation effect of these technologies can not always achieve the goal of high efficiency
Method used
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Embodiment 1
[0020] A method for promoting RNA degradation, inhibiting the expression of AMPKα gene in 293T cells. Specifically through the following methods:
[0021] 1) According to the sequence of SEQ ID NO.1, express and purify the fusion protein in prokaryotic;
[0022] 2) Add antisense oligonucleotides in Table 1 to the above-mentioned purified protein. The mixture is then added to the cultured cells;
[0023] Table 1 Antisense oligonucleotide sequence
[0024]
[0025] 3) After culturing for 48 hours again, the expression of AMPKα can be inhibited.
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The invention discloses a method for promoting RNA degradation; ribonuclease H is connected to a DNA binding structural domain by a genetic engineering method, and when a foreign DNA containing a specific recognition characteristic of the DNA binding structural domain is bound to a RNA, the endoribonuclease of the ribonuclease H of a fusion protein can actively degrade the RNA in a RNA / DNA hybridsystem. The method realizes the specific and efficient degradation of the nucleic acid molecule RNA, can be used for inhibiting gene expression and clearing RNA virus in vivo, and is simple and practical.
Description
technical field [0001] The invention belongs to the technical field of genetic engineering and relates to a method for promoting RNA degradation, specifically a method for degrading RNA in an RNA / DNA hybrid system based on ribonuclease H combined with a DNA binding domain. Background technique [0002] Degradation of target RNA has great application prospects: [0003] 1. Application in exploring gene function: In the post-genomic era, clarifying the biological role of functional gene expression products in the genome has far-reaching significance for the development of medicine. RNA degradation technology can quickly, economically and easily eliminate the expression of target genes in a sequence-specific manner, and has become an important research method for exploring gene functions. It is of great significance for elucidating signal transduction pathways and discovering new drug targets. [0004] 2. Application of RNA degradation technology in the field of gene therapy:...
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Login to View More IPC IPC(8): C07K19/00
CPCC07K2319/80C12N9/22C12Y301/26004
Inventor 李燕强
Owner 李燕强