46 results about "In vitro transfection" patented technology

The invention provides highly acidic chitosan-nucleic acid polyplex compositions. The compositions may be used to transfect cells in vitro and in vivo, and are particularly useful for transfecting cells of mucosal epithelia.

The invention provides a whole genome infectious clone of mink enteritis parvovirus and its construction method and application. The method is to digest the complete genome of the MEV replication form, and sequentially and directional clone it into the vector pUC18‑M in the form of segments to obtain the recombinant plasmid. After mixing the recombinant plasmid and the transfection reagent, the CRFK cells can be transfected to obtain the rescued virus. The mink enteritis parvovirus infectious clone constructed by using the reverse genetic technology in the present invention transfects CRFK cells in vitro, and can induce the cells to produce the same cytopathic and growth tendency as the parental virus. This method is simple, fast, time-saving and labor-saving, and has higher accuracy than the traditional PCR method. The application of this cloning system can quickly and conveniently carry out base mutations at any position in the MEV genome, thereby providing a basis for the subsequent development of MEV molecular biology. Research and the development of vaccines provide effective ways and means.

The invention provides a preparation method of a liposome-protected nano-gold gene carrier: dissolving dioctadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine in an organic solvent, and then removing the organic solvent After dissolving in water, ultrasonically obtain the phospholipid vesicle solution; the molar ratio of the dioctadecyldimethylammonium bromide and dioleoylphosphatidylethanolamine is 1:10~10:1; the phospholipid vesicle After the capsule solution is mixed with the compound containing gold ions, a reducing agent is added to react to obtain the nano-gold gene carrier protected by the liposome. After the liposome-protected nano-gold gene carrier of the present invention is combined with the gene, the transfection efficiency is high, and the gene release speed is fast in a specific environment; it has good stability in serum, no aggregation phenomenon, and does not need to be cultured during the in vitro transfection process. The serum in the liquid is removed, which simplifies the transfection process; the DNA load is high, which reduces the amount of DNA used, and reduces the cytotoxicity and immunogenicity of exogenous DNA to the body.

The invention discloses a degradable polymer based on ring opening polymerization of a valerolactone derivative and a production method and application thereof. The compound is mainly produced throughMichael reaction, ROP reaction and thio-ene click reaction, and the structure of the compound is identified through nuclear magnetism and mass spectrometry. According to the degradable polymer, through testing means of gel electrophoresis, dynamic light scattering, SEM and the like, the fact that the degradable polymer can condense DNA is proven, and the diameters of formed nanoparticles are allsmaller than 200 nm. An in-vitro transfection experiment proves that lipidosome formed by the degradable polymer and dioleoyl phosphatidylethanolamine (DOPE) can be used as a non-viral gene vector.