Abstract
Adenovirus (Ad) vectors are produced from molecular clones (MC) of the Ad genome. E1 and E3 domains are deleted; removal of E1 prevents virus replication. The genome is cloned into a plasmid vector. Infected cells provide viral RNA, for example, spike of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Spike sequences are amplified and cloned into a shuttle vector from where the expression cassette is excised and inserted into an Ad MC. Ad MC transfection of E1+ helper cells rescues the vaccine, which is expanded, tested, and ready for good manufacturing practice (GMP) production and clinical trials.
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