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And viral vectors can be produced by transfection with plasmids, stable packaging or producer cell lines, or infection using virus stocks. The large virus particles pose challenges during chromatography, and enveloped viruses are sensitive to processing conditions including shear force.

Typically, a viral vector is defined by three components as follows: (1) the protein capsid and/or envelope that encapsidates the genetic payload, and defines the vectorʼs tissue or cell tropism and antigen recognition; (2) the transgene of interest, which when expressed in cells, serves to confer a desired effect; and

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There are three basic ways to produce viral vectors: using a stable packaging cell line, using transient transfection, or using infection, Dr. van der Loo explains. For the latter, Iʼm referring to the baculovirus system developed by Robert Kotin when he was an investigator at the NIH.

In order to make vectors safe to use, all of the viral genes are removed, and the vector is modified to deliver only therapeutic genes. The choice of which viruses to use as vectors is based on three criteria: How well researchers understand the virus. How well the virus can target certain cell types.

Retroviral vectors are created by the removal of the retroviral gag, pol, and env genes, which are replaced by the therapeutic expression cassette. To produce vector particles, removed gag, pol, and env genes are provided in trans by specific packaging cell lines.