Semliki Forest virus and Sindbis virus vectors.
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.
Ehrengruber
Markus U
MU
University of Zurich, Zurich, Switzerland.
Lundstrom
Kenneth
K
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D005822:Genetic Vectors; D005826:Genetics, Medical; D006624:Hippocampus / Q000821:virology; D006801:Humans; D009474:Neurons / Q000821:virology; D010957:Plasmids / Q000235:genetics; D051381:Rats; D012093:Replicon; D012672:Semliki forest virus / Q000235:genetics* / Q000502:physiology; D012845:Sindbis Virus / Q000235:genetics* / Q000502:physiology; D046509:Tissue Culture Techniques; D014162:Transfection; D019065:Virus Assembly