Implantation of Isolated Carriers and receptors into living cells by Sendai virus envelope-mediated fusion

This chapter describes the implantation of isolated carriers and receptors into living cells by reconstituted sendai virus (RSVE) envelope-mediated fusion. RSVE can serve as an efficient carrier for the introduction of either soluble macromolecules or membrane components into eukaryotic cells. It is shown that when soluble macromolecules such as protein or DNA molecules are present in the detergent-solubilized fraction, they are trapped within the vesicle formed after removal of the detergent. Such loaded fusogenic vesicles have been used for the introduction of proteins and DNA molecules into living cultured cells. If a membrane component is added to the detergent-solubilized mixture of the viral glycoproteins and phospholipids, it is inserted into the viral envelope formed after removal of the detergent, resulting in the formation of hybrid fusogenic viral envelopes. Membranes rich in receptors for Epstein-Barr virus (EBV) from human lymphoma Raji cells are coreconstituted with RSVE. Fusion-mediated implantation of the EBV receptors into membranes of receptor-negative cells converted the latter susceptible to infection by EBV. Thus, it appears that RSVE may serve as a convenient and efficient vehicle for the introduction of ion channels as well as virus, lectin, or hormone receptors, or any other membrane components of well-defined structure or function into plasma membranes of living animal cells.