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Targeting exosome based on RBD region of SARS-CoV-2 S protein and preparation method of targeting exosome

A sars-cov-2s, targeted technology, applied in the field of biomedicine, can solve the problem of non-specific drugs or vaccines, and achieve the effect of inhibiting virus replication

Active Publication Date: 2021-01-15
SUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, no specific drug or vaccine has been officially approved

Method used

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  • Targeting exosome based on RBD region of SARS-CoV-2 S protein and preparation method of targeting exosome
  • Targeting exosome based on RBD region of SARS-CoV-2 S protein and preparation method of targeting exosome
  • Targeting exosome based on RBD region of SARS-CoV-2 S protein and preparation method of targeting exosome

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Embodiment 1

[0030] 1. Design primers for RBD (F: 5'-ATGTTTCCTAATATTACAAACTTGTGCC-3', SEQ ID NO.3; R: 5'-TTATGCTGGTGCATGTAGAAGTTCA-3', SEQ ID NO.4), using cDNA from throat swab samples from COVID-19 patients As a template, use the TaKaRa PCR kit to amplify the complete fragment of RBD, 2% agarose electrophoresis, Axygen gel cutting and recovery kit for DNA product purification; synthesize the full-length DNA of VSVG transmembrane region and intracellular region in vitro, and pass T4 enzyme The connection was inserted into the pCMV vector to construct the pCMV-VSVG vector, and the sequence was verified by sequencing;

[0031] 2. The RBD gene purified and recovered by PCR in step 1 was ligated with the pCMV-VSVG vector by T4 enzyme, and reacted overnight at 16°C to construct the pCMV-RBD-VSVG vector, which was verified by sequencing; through vector transformation, plating, and picking single clones , carry out expanded culture, and extract the pCMV-RBD-VSVG plasmid vector through the Axygen ...

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Abstract

The invention discloses a targeting exosome based on an RBD region of SARS-CoV-2 S protein and a preparation method of the targeting exosome, and belongs to the technical field of biological medicines. RBD-VSVG fusion protein is expressed on the targeting exosome, and the RBD-VSVG fusion protein is obtained by replacing an extracellular region of VSVG with RBD of SARS-CoV-2 S protein. According tothe invention, the targeting exosome capable of efficiently delivering anti-SARS-CoV-2 potential drugs in a tissue-specific manner is constructed; SARS-CoV-2 siRNA is wrapped by the targeting exosome, so that specific inhibition of virus replication in tissues and organs is realized. and in a mouse animal model, a tail vein injection exosome wraps the SARS-CoV-2 siRNA, so that the virus replication amount in mouse lung tissue can be remarkably inhibited, and pneumonia and other symptoms caused by virus infection are relieved.

Description

technical field [0001] The invention relates to a targeted exosome based on the RBD region of the SARS-CoV-2 S protein and a preparation method thereof, belonging to the technical field of biomedicine. Background technique [0002] Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of coronavirus disease 2019 (COVID-19), and its mortality rate has been rising, becoming a major global public health problem. So far, no specific drug or vaccine has been officially approved. Structural analysis and pathological observation confirmed that SARS-CoV-2 virus enters tissues and organs by binding to angiotensin-converting enzyme 2 (ACE-2) on host cells, and virus entry into cells depends on SARS-CoV-2 that specifically recognizes ACE2 The receptor binding domain (RBD) of the spike protein (S). Blocking RBD and ACE2 binding is currently considered to be a major potential strategy for the development of vaccines, neutralizing antibodies, and small molecule drugs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/85A61K47/46A61K47/42A61K45/00A61P31/14
CPCC12N5/0686C07K14/005C12N15/85A61K47/46A61K47/42A61K45/00A61P31/14C12N2510/00C07K2319/02C12N2770/20022C12N2760/20222C12N2800/107C12N2509/10C12N15/88C12N2310/14C12N15/1131C12N2320/32A01K2227/105A01K2217/072A01K2207/15A61K35/76Y02A50/30
Inventor 熊思东傅煜轩
Owner SUZHOU UNIV
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