title: Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters.
abstract: Transferring multiple genes into the same cell allows for the combination of genetic correction, marking, selection and conditional elimination of transduced cells or the reconstitution of multisubunit components and synergistic pathways. However, this cannot be reliably accomplished by current gene transfer technologies. Based on the finding that some cellular promoters intrinsically promote divergent transcription, we have developed synthetic bidirectional promoters that mediate coordinate transcription of two mRNAs in a ubiquitous or a tissue-specific manner. Lentiviral vectors incorporating the new promoters enabled efficient dual gene transfer in several tissues in vivo after direct delivery or transgenesis, and in a human gene therapy model. Because divergent gene pairs, likely transcribed from shared promoters, are common in the genome, the synthetic promoters that we developed may mimic a well-represented feature of transcription. Vectors incorporating these promoters should increase the power of gene function studies and expand the reach and safety of gene therapy.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, via Olgettina 58, 20132 Milano, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Amendola
authors.forename: Mary Anna
authors.initials: MA
authors.lastname: Venneri
authors.forename: Alessandra
authors.initials: A
authors.lastname: Biffi
authors.forename: Elisa
authors.initials: E
authors.lastname: Vigna
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
mesh_terms: D018952:Antigens, CD34 / Q000096:biosynthesis; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008156:Luciferases / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D019076:Transgenes
chemical_list: D018952:Antigens, CD34; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D008156:Luciferases