Abstract
Semliki Forest virus (SFV) and Sindbis virus (SIN) are two, positive-strand RNA viruses of the alphavirus genus. Vectors for both have been developed to express high levels of foreign genes in vitro and in vivo. Basic Protocol 1 describes the preparation of packaged SFV and SIN replicons by co-electroporation of helper and vector RNA into baby hamster kidney (BHK)-21 cells. Basic Protocol 2 describes the activation of packaged SFV replicons with a-chymotrypsin. Basic Protocol 3 provides a method for the infection of hippocampal slices. Basic Protocol 4 is a technique for the infection of primary cultures of dispersed neurons with infectious SFV and SIN replicons. The Alternate Protocol describes a method for the cotransfection of in vitro-transcribed vector and helper RNA into BHK-21 cells. Support Protocol 1 describes determining the titers of infectious SFV and SIN replicon stocks, and Support Protocol 2 for metabolic labeling of infected cells.
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