AAV vector based generic gene therapy drug for coronavirus infection and preparation method thereof

Abstract

The invention relates to an AAV vector based generic gene therapy drug for coronavirus infection. The therapy drug includes adeno-associated viruses; the adeno-associated viruses include AAV capsid, ssAAV genomes packed in the AAV capsid, promoters and sequences suitable for coding soluble extracellular region ACE2; and the sequence-encoded protein can be secreted out of cells and has a functionaldomain specifically binding to S protein. The gene sequence encoded with the soluble extracellular region ACE2 protein is introduced into target cells by using the AAV vector, so that the target cells can secret the soluble ACE2 to the outside of the cells so as to bind to the S protein of coronavirus, and therefore, the purpose of antagonizing SARS-CoV infection can be achieved. The gene sequence is the gene fragments encoding the soluble extracellular region ACE2 (1-740) and (1-620) non-full-length protein obtained through deletion based on the gene sequence encoding ACE2 full length protein (1-805), and both the two non-full-length protein reserve the function domain specifically binding to the S protein, can be secreted out of the cells and used for inhibiting the infection of the novel coronavirus.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular to a general-purpose gene therapy drug for coronavirus infection based on AAV vectors. Background technique [0002] The World Health Organization officially named the disease caused by SARS-CoV-2 as COVID-19 (coronavirusdisease-2019). About 25% of people infected with COVID-19 are severely ill. The typical feature is severe viral pneumonia, which is extremely severe and life-threatening. The crude case fatality rate is about 0.5%-1%. [0003] SARS-CoV-2 is similar to SARS and has similar genome composition and virus structure characteristics. Both of them recognize and bind ACE2 (angiotensin-converting enzyme 2) on the surface of target cells through the RBD (receptor binding domain) of Spike protein. Body, through membrane fusion, enters the endosome / lysosome pathway, uncapsids, and releases viral genome RNA. In the cytoplasm, the full-length positive-strand genome of t...

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Problems solved by technology

However, the natural AAV genome is single-stranded DNA. For a single-stranded AAV (Adeno-associated Virus, AAV) vector, its capacity is 4.7kb, which can accommodate promoters, soluble ACE2 coding regions and Poly A sequences, but its disadvantages It takes a certain time for the single-chain AAV vector to convert into a double-chain after entering the cell, which makes the expression of soluble ACE2 lag behind

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