Highly efficient baculovirus-mediated multigene delivery in primary cells

Maysam Mansouri, Itxaso Bellon-Echeverria, Aurélien Rizk, Zahra Ehsaei, Chiara Cianciolo Cosentino, Catarina S. Silva, Ye Xie, Frederick M. Boyce, M. Wayne Davis, Stephan C. F. Neuhauss, Verdon Taylor, Kurt Ballmer-Hofer, Imre Berger () and Philipp Berger ()
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Maysam Mansouri: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute
Itxaso Bellon-Echeverria: European Molecular Biology Laboratory (EMBL)
Aurélien Rizk: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute
Zahra Ehsaei: University of Basel
Chiara Cianciolo Cosentino: Institute of Molecular Life Sciences, University of Zürich
Catarina S. Silva: European Molecular Biology Laboratory (EMBL)
Ye Xie: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute
Frederick M. Boyce: Massachusetts General Hospital
M. Wayne Davis: University of Utah
Stephan C. F. Neuhauss: Institute of Molecular Life Sciences, University of Zürich
Verdon Taylor: University of Basel
Kurt Ballmer-Hofer: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute
Imre Berger: European Molecular Biology Laboratory (EMBL)
Philipp Berger: Biomolecular Research, Molecular Cell Biology, Paul Scherrer Institute

Nature Communications, 2016, vol. 7, issue 1, 1-13

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11529

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