title: Protein transfer into human cells by VSV-G-induced nanovesicles.
abstract: Identification of new techniques to express proteins into mammal cells is of particular interest for both research and medical purposes. The present study describes the use of engineered vesicles to deliver exogenous proteins into human cells. We show that overexpression of the spike glycoprotein of the vesicular stomatitis virus (VSV-G) in human cells induces the release of fusogenic vesicles named gesicles. Biochemical and functional studies revealed that gesicles incorporated proteins from producer cells and could deliver them to recipient cells. This protein-transduction method allows the direct transport of cytoplasmic, nuclear or surface proteins in target cells. This was demonstrated by showing that the TetR transactivator and the receptor for the murine leukemia virus (MLV) envelope [murine cationic amino acid transporter-1 (mCAT-1)] were efficiently delivered by gesicles in various cell types. We further shows that gesicle-mediated transfer of mCAT-1 confers to human fibroblasts a robust permissiveness to ecotropic vectors, allowing the generation of human-induced pluripotent stem cells in level 2 biosafety facilities. This highlights the great potential of mCAT-1 gesicles to increase the safety of experiments using retro/lentivectors. Besides this, gesicles is a versatile tool highly valuable for the nongenetic delivery of functions such as transcription factors or genome engineering agents.
authors.affiliation: Université de Lyon, INSERM, U851, Lyon, France. philippe.mangeot@inserm.fr
authors.forename: Philippe-Emmanuel
authors.initials: PE
authors.lastname: Mangeot
authors.forename: Sandra
authors.initials: S
authors.lastname: Dollet
authors.forename: Mathilde
authors.initials: M
authors.lastname: Girard
authors.forename: Claire
authors.initials: C
authors.lastname: Ciancia
authors.forename: Stéphane
authors.initials: S
authors.lastname: Joly
authors.forename: Marc
authors.initials: M
authors.lastname: Peschanski
authors.forename: Vincent
authors.initials: V
authors.lastname: Lotteau
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D019295:Computational Biology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D009052:Leukemia Virus, Murine / Q000235:genetics; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission; D039904:Pluripotent Stem Cells / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014161:Transduction, Genetic; D018116:Vesiculovirus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D050916:TRPV Cation Channels; D014157:Transcription Factors; C446923:Trpv6 protein, mouse; D014759:Viral Envelope Proteins