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Gene targeting novel coronavirus spinous process, kit, screening method and application

A coronavirus, screening method technology, applied in the field of genetic engineering

Active Publication Date: 2021-01-01
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Abstract
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Problems solved by technology

[0017] In order to solve the problem that the existing drugs cannot specifically target the spine (S), the key structure of SARS-CoV-2, the present invention combines bioinformatics methods and RNA editing technology based on the CRISPR-Cas13a system, and proposes a CRISPR-Cas13a-based The technology of Cas13a system to recognize and degrade SARS-CoV-2 S in human cells

Method used

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  • Gene targeting novel coronavirus spinous process, kit, screening method and application
  • Gene targeting novel coronavirus spinous process, kit, screening method and application
  • Gene targeting novel coronavirus spinous process, kit, screening method and application

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Embodiment

[0082] 1. Search for specific fragments of SARS-CoV-2 S: using comprehensive methods such as sequence alignment, three-dimensional structure superposition analysis, and molecular docking simulation. Such as figure 1 The provided amino acid sequence alignment and three-dimensional structure overlay alignment of the receptor binding domain (RBD) of SARS-CoV-2 S and SARS-CoV S.

[0083] (1) Sequence alignment: from NCBI database ( https: / / www.ncbi.nlm.nih.gov / ) for SARS-CoV-2 S, gene sequences of SARS-CoV S and MERS-CoV S. The sequence numbers of SARS-CoV-2 S, SARS-CoV S and MERS-CoV S are MN908947.3, AY278488.2 and KF600652.1, respectively. S-protein sequence alignment and visualization was performed using Clustal w2.1 and Exprint 2.0.

[0084] According to the sequence alignment results, the receptor-binding domain (Receptor-Binding-Domain, RBM) of the SARS-CoV-2 S protein has two residues that are significantly different from the residues of the corresponding fragment of t...

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Abstract

The invention discloses a gene targeting novel coronavirus spinous process, a kit, a screening method and application, and relates to the technical field of gene engineering. The RNA sequence of the gene for targeting the novel coronavirus SARS-CoV-2 spinous process is shown as SEQ ID NO: 6. The specific treatment kit for resisting the SARS-CoV-2 infection comprises the gene. According to the gene, the crRNA sequence with the highest S degradation efficiency is screened out, so that the purpose of effectively reducing the virus load in cells is achieved. According to the gene, bioinformatics simulation and biological experiments are ingeniously combined, and the CRISPR-Cas13a system designed in a targeted mode can specifically recognize and degrade S RNA in human alveolar epithelial cells.The crRNA designed by the invention is expected to be applied to development of specific treatment tools and medicines for resisting SARS-CoV-2 infection.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a gene targeting novel coronavirus spines, a kit, a screening method and an application. Background technique [0002] Currently, the novel coronavirus SARS-CoV-2 has caused the global pandemic of coronavirus pneumonia (COVID-19), but there is currently no specific therapeutic drug against SARS-CoV-2. [0003] Although antiviral drugs such as lopinavir, ritonavir, remdesivir and chloroquine have shown inhibitory activity against SARS-CoV or SARS-CoV-2 in vitro, the clinical outcomes of these drugs in patients with COVID-19 No apparent benefit. [0004] However, clinical evidence does not recommend the use of corticosteroids for treatment because of its limited benefit and serious complications. Convalescent plasma or immune globulin has potential therapeutic capabilities, but its safety and efficacy still need to be verified by large-scale clinical trial...

Claims

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Application Information

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IPC IPC(8): C12N15/50C12N15/867C12N9/22C12N15/113C07K14/165C40B40/06C40B50/06A61K31/7088A61P31/14
CPCC07K14/005C12N9/22C12N15/86C12N15/113C40B40/06C40B50/06A61K31/7088A61P31/14C12N2310/20C12N2740/15043C12N2770/20022
Inventor 康春生王琳周俊虎王琦雪王蕴非
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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