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Method for screening siRNA on basis of bioinformatics

A bioinformatics and gene technology, applied in the field of small nucleic acid drugs, to achieve the effect of avoiding spread, avoiding toxic and side effects, and rapid response

Inactive Publication Date: 2015-03-18
HANGZHOU CONVERD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, although many small nucleic acid drugs have entered preclinical research in the research and development of drugs for the prevention and treatment of major diseases such as tumors, viruses, and cardiovascular diseases, they are still difficult to achieve in terms of drug delivery, targeting, industrial production scale and cost, and human safety. Bottleneck problems still exist, and these problems are more prominent in the field of emergency drug research and development for viral infections and infectious diseases. Therefore, it is necessary to develop new ideas, establish new technologies and new approaches to solve the above bottleneck problems, so as to speed up the pace of its industrialization, especially to improve the efficiency of new emergency drugs. "Gene-to-Drug" Efficiency in R&D

Method used

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Experimental program
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Effect test

Embodiment 1

[0087] The cRNA sequence (GeneBank accession number: KC853766.1) of a H7N9 influenza A virus genome HA protein was obtained from NCBI (National Center for Biotechnology Information, website: www.ncbi.nlm.nih.gov), with a length of 1689bp.

[0088] Using the cRNA sequence as a template, use the siDirect online design tool to design the first candidate siRNA sequence. Specifically, the cRNA sequence was input into the siDirect online design tool (URL: http: / / sidirect2.rnai.jp / ), and 67 first candidate siRNA sequences were obtained.

[0089] The obtained first candidate siRNA sequences were sorted according to the thermodynamic stability of the double helix formed by the seed region and the target sequence (seed duplex stability), and the highest stability (T m value ≤ 10°C), the first candidate siRNA sequence has 4 corresponding target site sequences:

[0090] Sequence 1: 5'-ACCAAAGTAAACACATTAACTGA-3'

[0091] Sequence 2: 5'-TAGAATTTTCAGCCGATTTAATT-3'

[0092] Sequence 3: 5'-...

Embodiment 2

[0100] The cRNA sequence (GeneBank accession number: KC853766.1) of a H7N9 influenza A virus genome HA protein was obtained from NCBI (National Center for Biotechnology Information, website: www.ncbi.nlm.nih.gov / ), with a length of 1689bp .

[0101] Using the cRNA sequence as a template, the first candidate siRNA sequence was designed using siDirect, Dharmacon, and GenScript online design tools, respectively. Specifically, the cRNA sequence was input into (1) siDirect online design tool (URL: http: / / sidirect2.rnai.jp / ), (2) Dharmacon online design tool (URL: http: / / www.thermoscientificbio.com / design-center / ?redirect=true) and (3) GenScript online design tool (URL: https: / / www.genscript.com / ssl-bin / app / rnai), get the first candidate siRNA sequence and The ranking results are compared and those first candidate siRNA sequences with higher comprehensive scores are selected.

[0102] Eliminate the first candidate siRNA sequence that can produce off-target effects on human genes ...

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Abstract

The invention provides a method for screening siRNA on basis of bioinformatics. The siRNA obtained through the method is capable of silencing or suppressing the human respiratory virus genes without silencing or suppressing the human genes. The method comprises the following steps: 1) providing a complete genome sequence of the human respiratory viruses; 2) designing first candidate siRNA sequences by taking the complete genome sequence of the human respiratory viruses as a template, and ensuring that the first candidate siRNA sequences are capable of silencing or suppressing the human respiratory virus genes; and 3) excluding the first candidate siRNA sequences which are capable of generating off-target effect to the human genes through comparative analysis so as to obtain second siRNA sequences which do not silence or suppress the human genes. According to the method, rapid response can be carried out in allusion to the new burst respiratory virus epidemic situations, so that the spreading of the acute epidemic situations can be avoided; and the siRNA type drugs obtained through the method are suitable for mucosal immunity.

Description

technical field [0001] The present invention relates to the field of small nucleic acid medicines, in particular to a bioinformatics-based screening method for siRNA against respiratory viruses, siRNA obtained by the method, pharmaceutical composition, use, and device. Background technique [0002] Respiratory viruses refer to viruses that can invade the respiratory tract and cause local lesions of the respiratory tract or only use the respiratory tract as an invasion portal, mainly causing lesions of tissues and organs outside the respiratory tract. Respiratory viruses are mostly RNA viruses. When RNA viruses multiply rapidly in living cells (genome replication and virion assembly), they can easily change their genetic structure in different tissues in a short period of time, with strong variability. [0003] Among respiratory viruses, influenza virus (influenza virus, referred to as influenza virus for short), especially influenza A virus, is the most important in causing ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/113C12M1/00A61K48/00A61P31/14
Inventor 胡放赵荣华张庆勇李树龙孙言伟
Owner HANGZHOU CONVERD CO LTD
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