Highly efficient baculovirus-mediated multigene delivery in primary cells

作者：Maysam Mansouri, Itxaso Bellón-Echeverría, Aurélien Rizk, Zahra Ehsaei, Chiara 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 · 发表于：Nature Communications · 年份：2016 · DOI：10.1038/ncomms11529 · 被引用次数：109 · 研究领域：Viral Infectious Diseases and Gene Expression in Insects、CRISPR and Genetic Engineering、Pluripotent Stem Cells Research

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.