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Medicine capable of simultaneously targeting RNA (Ribonucleic Acid) and DNA (Deoxyribonucleic Acid) viruses and application

A drug and virus technology, applied in the field of biomedicine, to achieve the effect of small molecular weight, simple synthesis, small economy and time cost

Pending Publication Date: 2022-05-27
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no nucleic acid-based antiviral drug that can fight not only DNA viruses but also ssRNA viruses without the need for DNA intermediates

Method used

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  • Medicine capable of simultaneously targeting RNA (Ribonucleic Acid) and DNA (Deoxyribonucleic Acid) viruses and application
  • Medicine capable of simultaneously targeting RNA (Ribonucleic Acid) and DNA (Deoxyribonucleic Acid) viruses and application
  • Medicine capable of simultaneously targeting RNA (Ribonucleic Acid) and DNA (Deoxyribonucleic Acid) viruses and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 AntiV-SGN can inhibit the expression level of SARS-CoV-2 reporter plasmid in mammalian cells

[0057] The ssRNA genome of the RNA virus SARS-CoV-2 is 30,000-nt and contains 12 coding functional open reading frames. According to previous studies, the end of its genome is the nucleocapsid protein (N) gene, which is a highly conserved region that encodes the capsid protein for viral packaging. Since our AntiV-SGN system has no target sequence restriction (such as PAM / PFS), theoretically any position in the N gene can be a candidate position of hpDNA. We ruled out leader sequences that might bind in the human transcriptome, designed and synthesized four hpDNA targeting N genes as figure 2 - A (Hp-CoV-1, Hp-CoV-2, Hp-CoV-3, Hp-CoV-4).

[0058] In order to verify whether AntiV-SGN (nucleotide sequence as shown in SEQ ID NO.1, encoded amino acid sequence as shown in SEQID NO.2) can successfully inhibit RNA virus SARS-CoV-2 reporter plasmid in mammalian cells For ...

Embodiment 2

[0070] Example 2 AntiV-SGN can inhibit the expression level of HBV reporter plasmid in mammalian cells

[0071] In order to verify whether AntiV-SGN can successfully inhibit the expression of DNA virus HBV reporter plasmid in mammalian cells, we synthesized a gene encoding A. fulgidus FEN1 protein (1011bp, 336 amino acids, 35kDa) and linked An NLS for optimal expression in the nucleus NLS-FEN1 plasmid [1] .

[0072] Then construct the HBV reporter plasmid, first use restriction endonuclease HindIII and KpnI to carry out digestion, pEGFP-N1 plasmid is linearized, secondly obtain the DNA fragment containing X protein sequence by PCR amplification and use restriction endonuclease HindIII and KpnI KpnI was digested to obtain DNA fragments with cohesive ends (primers were HBx-F-HindIII and HBx-R-KpnI, respectively), and finally the linearized vector was enzyme-ligated with the DNA fragment containing the X protein sequence by T4 DNA ligase to obtain HBV reporter plasmid (the targ...

Embodiment 3

[0077] Example 3 Comparison of AntiV-SGN with existing gene editing tools RNAi and CRISPR

[0078] Both CRISPR system and RNAi are widely used in gene editing technology. Among them, the CRISPR system has always had the problem of inconvenient delivery in vivo due to the relatively large size of the Cas protein (1367 amino acids to 422 amino acids). Although some recent studies have reported that RNA-targeting Cas9 (RCas9) can be used to edit RNA, however, in specific In research, Cas9 is usually used to edit DNA viruses, and Cas13 is usually used to edit RNA viruses. While the AntiV-SGN system only needs to change the leader sequence (20-nt) of hpDNAs when targeting different subtypes of different viruses, it does not need to change the protein, making AntiV-SGN an ideal supplement to traditional antiviral drugs in low cost and time.

[0079] In order to prove the advantages of the AntiV-SGN system, we first compared its differences with CRISPR-Cas13 and siRNA in HEK293, A54...

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Abstract

The invention discloses a drug for simultaneously targeting RNA and DNA viruses and application. The drug is a drug for broad-spectrum antiviral gene editing. The drug comprises an AntiV-SGN system, and the AntiV-SGN system comprises (i) at least one oligonucleotide probe and (ii) an AntiV-SGN protein molecule or a polynucleotide encoding the protein molecule. The AntiV-SGN system can be used for destroying nucleic acid of DNA viruses and can also act on destroying nucleic acid of RNA viruses, and it is verified that the AntiV-SGN system can inhibit virus replication and expression in mammalian cells, so that virus infection is inhibited; the system is high in universality, small in size and simple in component, in-vivo delivery is facilitated, significant exploration is performed on solving the problems occurring in development of existing antiviral drugs, and the system has good clinical application potential.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a medicine for fighting viruses and its application. Background technique [0002] Throughout history, human beings have encountered many different types of unknown and sudden virus attacks several times, but they lack powerful anti-"weapons". Moreover, only a few known virus vaccines and antiviral drugs have been approved for marketing. We still lack powerful means to deal with many sudden attacks of unknown viruses. [0003] So far, the control and treatment of viruses mainly rely on vaccines, small molecule drugs, traditional Chinese medicine, antibodies and so on. Among them, it takes several years to develop vaccines and antibodies. Small molecule drugs inhibit virus replication by activating host cells to synthesize anti-viral functional proteins or proteins that inhibit viral DNA transcription activity. These antiviral approaches require a comprehensive u...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/46C12N9/16C12N15/11A61P31/14A61P31/20
CPCC12N9/16C12Y301/00A61K48/005A61K48/0025A61K38/465A61P31/20A61P31/14Y02A50/30
Inventor 徐澍田坤张赟郭永健
Owner CHINA PHARM UNIV
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