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  • Highly efficient baculovirus-mediated multigene delivery in primary cells.

Highly efficient baculovirus-mediated multigene delivery in primary cells.

Nature communications (2016-05-05)
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, Philipp Berger
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
Anti-β-Tubulin III (neuronal) antibody, Mouse monoclonal, ~1.0 mg/mL, clone 2G10, purified from hybridoma cell culture