8 results about "Slow virus" patented technology

This invention provides a method for constructing and applying MDCK cells stably integrated with disialotyltransferase, relating to the field of biotechnology. The construction method includes: infecting MDCK cells with lentiviral particles, followed by culture and screening to obtain MDCK cells stably expressing porcine ST3GAL1 and porcine ST6GAL1 genes; the lentiviral particles carry porcine ST3GAL1 and porcine ST6GAL1 genes; the nucleic acid sequence of the porcine ST3GAL1 gene is shown in SEQ ID NO.1; the nucleic acid sequence of the porcine ST6GAL1 gene is shown in SEQ ID NO.2. This construction method is simple and convenient, and the constructed MDCK cells can stably express porcine ST3GAL1 and porcine ST6GAL1 genes, effectively improving the culture efficiency of swine influenza virus, and thus can be used in the preparation of swine influenza virus.

Formulas for use in enhancing the transduction efficiency of lentiviral vector (LV) particles into various target cells are disclosed herein. For example, a formulation for use in enhancing the transduction of paramyxovirus pseudotype lentiviral vector particles into target cells may comprise a cell-permeable peptide (CPP) comprising at least one of Vectofusion-1 or LAH4-C; and poloxamer 407, wherein the concentration of poloxamer 407 is greater than or equal to the concentration of CPP. Additionally, a method of using such formulations to enhance the transduction of LV particles, including particularly in vivo transduction, is disclosed herein.

This invention discloses a nucleic acid molecule encoding a modified retroviral Gag protein and its applications. The Gag protein contains an engineered acylated tag sequence at its N-terminus, comprising: a glycine residue at the second position after the initiating methionine for myristylation modification; and / or a cysteine ​​residue in the N-terminal region for palmitoylation modification. The tag sequence is selected from SEQ ID NO:1, 3-9, or derivative amino acid sequences having at least 80% identity and retaining function. Preferably, the modification involves replacing the first 9 amino acids at the N-terminus of the wild-type HIV-1 Gag protein with the aforementioned tag to enhance membrane anchoring and improve viral packaging efficiency. This invention also provides packaging plasmids containing this molecule, host cells, packaging systems, and preparation methods. This invention can significantly improve viral yield and titer, and is particularly suitable for overcoming the efficiency bottleneck in packaging large-load lentiviruses of 7-10 kb.

The application belongs to the technical field of biological medicine, and discloses a spinal cord injury treatment drug based on a LINC00311 / miR-4318 / MYLK regulation axis and an application thereof, and aims to solve the technical problems of poor targeting and large side effects of the existing anti-inflammatory treatment of spinal cord injury. The effective component of the drug is an inhibitor of LINC00311, the inhibitor is selected from one or more of shRNA, siRNA, antisense oligonucleotide and CRISPRi system, can target the specific nucleotide sequence of LINC00311, and can be used in combination with miR-4318 mimic, and is administered by intrathecal injection, slow virus carrier delivery and the like. The in-vitro experiment proves that the inhibitor can significantly inhibit the release of TNF-alpha, IL-1beta, IL-6 and other pro-inflammatory factors by microglia cells; the in-vivo experiment shows that it can significantly improve the BBB motor function score of spinal cord injury rats, and reduce spinal cord tissue edema and necrosis. The application first discloses the mechanism of the LINC00311 / miR-4318 / MYLK ceRNA regulation axis in spinal cord injury, has the advantages of novel target, significant effect and high transformation potential, and provides a new scheme for the clinical treatment of spinal cord injury.

This invention provides a method for characterizing recombinant viral particle preparations using analytical ultracentrifugation. The recombinant viral particles include recombinant adeno-associated viral particles, recombinant adenovirus particles, recombinant lentivirus particles, and recombinant herpes simplex virus particles. Different variants of recombinant viral particles, including empty capsids and recombinant viral particles with different genomes (e.g., truncated genomes, aggregates, recombinants), can be identified and quantified. This method can be used to characterize recombinant viral particle preparations regardless of the genomic sequence of the recombinant virus or the serotype of the recombinant viral capsid.

An engineered enucleated mesenchymal stem cell (EMC) and its preparation method and application are disclosed, which can effectively solve the problem of preparing engineered enucleated EMC and realize its application in the preparation of drugs for treating herpesvirus infections. The steps are as follows: Lentiviral transfection is used to obtain EMC overexpressing Nectin-1, the overexpression being achieved through genetic engineering, including transfection of EMC using a lentiviral vector or adenovirus vector; then, enucleation is performed using discontinuous density gradient centrifugation to obtain the engineered enucleated EMC. The engineered enucleated EMC of this invention acts as "bait" to competitively capture viruses; the enucleation design fundamentally eliminates the risk of viral replication and leakage within cells; and the captured viruses are subsequently cleared along with the engineered enucleated EMC's own programmed apoptosis and subsequent macrophage phagocytosis, thereby achieving safe and efficient virus capture and physical clearance.

The application relates to the technical field of the cross of microfluidic technology, chemical synthesis and biomedicine, and discloses a method for preparing EGFR CAR-NK cells based on polypeptide gel droplet confined transfection technology and application, which comprises the following steps: (1) preparing collagen-mimic-peptide-modified alginate; (2) constructing a confined transfection system: adopting a droplet microfluidic chip, wrapping NK cells and slow virus carrying an EGFR CAR gene in the collagen-mimic-peptide-modified alginate droplet prepared in the step (1), and obtaining a gel droplet after gelation of the droplet; and (3) confined transfection and cell culture. The method can significantly reduce the dosage of slow virus and cell toxicity, greatly improve the transfection success rate of the CAR gene, the survival rate and functional activity of the NK cells, and finally obtain EGFR CAR-NK cell products with higher purity and stronger target killing capacity.