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Based on the research philosophy of translational medicine, Ruisai knows that the cause or appearance of each clinical disease is initially caused by a mutation in a certain gene in the individual. Therefore, for each clinical disease study, the initial source of the overall project project must also start with a gene.
In many years of translational medicine research, Ruisai is in the overall project cooperation of various clinicians. It is not only the overall thinking of the clinical research topics of translational medicine, but also the general experimental direction, and the operation summary of each specific experiment in each link.
The general experimental scheme of the whole experiment and the experimental methods of each individual technology are shared here, not only for the needs of the laboratory, but also to further improve the efficiency of translational medicine and to meet the needs of patients.
General experimental ideas for the overall project:
Gene-gene chemical synthesis/acquisition-vector construction-virus packaging-stabilized strain construction-cell function experiment-animal experiment/clinical sample analysis-data analysis statistics, research
Virus Packaging Technology 1 - Virus and Non-viral Vectors
Advantages of viral vectors
Characteristics
The viral vector is derived from the manipulation and modification of the viral genome, eliminating the pathogenic gene and retaining its infectious activity, making it a biologically active gene vector. It carries a foreign gene and is packaged into viral particles constitute a gene delivery system (gene delivery system).
Due to the diversity of the virus and the body's complex dependencies with people still understanding the life cycle of many viruses, molecular biology, such as the relationship between the occurrence and development of diseases and is far from comprehensive, it has a very technically transformation The big limitation is that not all virus types can be modified to eliminate their pathogenicity for the human body . In the past 20 years, several viruses such as retroviruses (including HIV), adenoviruses, adeno-associated viruses, herpes viruses (including herpes simplex virus, vaccinia virus and Epstein-Barr virus) have been successfully transformed into gene transfer vectors and carried out. Different levels of application. However , the development of molecular virology and biotechnology has applied reverse genetics methods to the construction of viral vectors, such as RNA viruses such as Sindbis virus, influenza virus, Sendai virus, etc., which have been successfully transformed into viral vectors.
Viral vectors for gene introduction and clinical research are generally characterized
(1) carrying foreign genes and capable of being packaged into viral particles;
(2) It can infect the target cells and mediate gene expression ;
(3) It does not cause disease to the human body and cannot spread and proliferate in the general environment.
Non-viral vector
Non-viral vectors include plasmid DNA , antisense oligonucleotides, Ribozyme, siRNA, and the like. Substances commonly used in the composition of non-viral vector introduction systems include lipids, polysaccharides, polypeptides or proteins, polyhistidine or polylysine, polyvalent cationic compounds, steroids and the like. Combinations of these different materials and can be encapsulated in a ratio to construct vector DNA or other nucleic acid material, such "artificial casing" mimic structural components constituting the liposome membrane skeleton. Common ingredients are liposomes, chitosan, polyvalent cationic polymers such as PEI, polylysine or histidine , polycationic compounds . At the same time plus a targeting polypeptide, polypeptide drugs derived from cell membrane fusion, a peptide fragment of the influenza virus HA protein to help targeting vector introduced, fusion, etc. released from the body endocytosis. However, the current non-viral vector introduction system is still not ideal in terms of transduction efficiency, particle uniformity, stability, etc., thus limiting the space for further development .
