title: Highly efficient baculovirus-mediated multigene delivery in primary cells.
abstract: Multigene delivery and subsequent cellular expression is emerging as a key technology required in diverse research fields including, synthetic and structural biology, cellular reprogramming and functional pharmaceutical screening. Current viral delivery systems such as retro- and adenoviruses suffer from limited DNA cargo capacity, thus impeding unrestricted multigene expression. We developed MultiPrime, a modular, non-cytotoxic, non-integrating, baculovirus-based vector system expediting highly efficient transient multigene expression from a variety of promoters. MultiPrime viruses efficiently transduce a wide range of cell types, including non-dividing primary neurons and induced-pluripotent stem cells (iPS). We show that MultiPrime can be used for reprogramming, and for genome editing and engineering by CRISPR/Cas9. Moreover, we implemented dual-host-specific cassettes enabling multiprotein expression in insect and mammalian cells using a single reagent. Our experiments establish MultiPrime as a powerful and highly efficient tool, to deliver multiple genes for a wide range of applications in primary and established mammalian cells.
authors.affiliation: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Maysam
authors.initials: M
authors.lastname: Mansouri
authors.affiliation: European Molecular Biology Laboratory (EMBL), Grenoble Outstation, B.P. 181, 38042 Grenoble Cedex 9, France.
authors.forename: Itxaso
authors.initials: I
authors.lastname: Bellon-Echeverria
authors.affiliation: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Rizk
authors.affiliation: Department of Biomedicine, University of Basel, CH-4058 Basel, Switzerland.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Ehsaei
authors.affiliation: Institute of Molecular Life Sciences, University of Zürich, CH-8057 Zürich, Switzerland.
authors.forename: Chiara
authors.initials: C
authors.lastname: Cianciolo Cosentino
authors.affiliation: European Molecular Biology Laboratory (EMBL), Grenoble Outstation, B.P. 181, 38042 Grenoble Cedex 9, France.
authors.forename: Catarina S
authors.initials: CS
authors.lastname: Silva
authors.affiliation: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Ye
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Department of Neurology, Massachusetts General Hospital, Cambridge, Massachusetts 02139, USA.
authors.forename: Frederick M
authors.initials: FM
authors.lastname: Boyce
authors.affiliation: Department of Biology and Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112-0840, USA.
authors.forename: M Wayne
authors.initials: MW
authors.lastname: Davis
authors.affiliation: Institute of Molecular Life Sciences, University of Zürich, CH-8057 Zürich, Switzerland.
authors.forename: Stephan C F
authors.initials: SC
authors.lastname: Neuhauss
authors.affiliation: Department of Biomedicine, University of Basel, CH-4058 Basel, Switzerland.
authors.forename: Verdon
authors.initials: V
authors.lastname: Taylor
authors.affiliation: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Kurt
authors.initials: K
authors.lastname: Ballmer-Hofer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Grenoble Outstation, B.P. 181, 38042 Grenoble Cedex 9, France.|School of Biochemistry, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
authors.affiliation: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Philipp
authors.initials: P
authors.lastname: Berger
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D019556:COS Cells; D064113:CRISPR-Cas Systems / Q000235:genetics; D002478:Cells, Cultured; D000067470:Cellular Reprogramming Techniques / Q000379:methods; D002522:Chlorocebus aethiops; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D061987:Sf9 Cells; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics