Highly efficient baculovirus-mediated multigene delivery in primary cells
| Autor: | Mansouri M, Itxaso Bellon-Echeverria, Rizk A, Ehsaei Z, Cianciolo Cosentino C, Cs, Silva, Xie Y, Fm, Boyce, Mw, Davis, Sc, Neuhauss, Taylor V, Ballmer-Hofer K, Berger I, Berger P |
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| Přispěvatelé: | Paul Scherrer Institute (PSI), European Molecular Biology Laboratory [Grenoble] (EMBL), University of Basel, Department of Biomedicine, University of Basel (Unibas), Institute of Molecular Life Sciences, University of Zurich, Universität Zürich [Zürich] = University of Zurich (UZH), Boston University School of Medicine (BUSM), Boston University [Boston] (BU), University of Utah, Department of Biology, Swiss National Science Foundation (31003A-146975 and 3100A0-118351), European Project: 279039,EC:FP7:HEALTH,FP7-HEALTH-2011-two-stage,COMPLEXINC(2011), European Project: 613879,EC:FP7:KBBE,FP7-KBBE-2013-7-single-stage,SYNSIGNAL(2013), European Project: 283570,EC:FP7:INFRA,FP7-INFRASTRUCTURES-2011-1,BIOSTRUCT-X(2011), University of Zurich, Berger, Imre |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Genome engineering [SDV]Life Sciences [q-bio] General Physics and Astronomy Primary cells Synthetic biology Genome editing Chlorocebus aethiops Sf9 Cells CRISPR Cellular Reprogramming Techniques Transgenes MultiPrime Cells Cultured ComputingMilieux_MISCELLANEOUS Genetics Multidisciplinary Gene Transfer Techniques 10124 Institute of Molecular Life Sciences 3100 General Physics and Astronomy Multigene expression COS Cells Mammalia Baculoviridae Reprogramming Recombinant baculoviral genomes Science Genetic Vectors 1600 General Chemistry Computational biology Biology Transfection Article General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 1300 General Biochemistry Genetics and Molecular Biology Animals Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology CRISPR/Cas9 Cas9 Bristol BioDesign Institute HEK 293 cells Baculovirus based expression systems General Chemistry Cell cultures SYNTHETIC BIOLOGY HEK293 Cells 030104 developmental biology Structural biology 570 Life sciences biology Gene expression CRISPR-Cas Systems Insect HeLa Cells |
| Zdroj: | Nature Communications Nature Communications, Nature Publishing Group, 2016, 7, pp.11529 Europe PubMed Central Maysam Mansouri, M, Bellon-Echeverria, I, Rizk, A, Ehsaei, Z, Cianciolo Cosentino, C, Silva, C S & Berger, I 2016, ' Highly efficient baculovirus-mediated multigene delivery in primary cells ', Nature Communications, vol. 7, 11529 (2016) . https://doi.org/10.1038/ncomms11529 Nature communications Nature Communications, Vol 7, Iss 1, Pp 1-13 (2016) |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/ncomms11529 |
| Popis: | 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. Current viral gene delivery systems are limited in the amount of foreign DNA they can deliver to cells. Here the authors develop MultiPrime, a baculovirus-based vector system capable of multigene delivery into a wide variety of cells, and use Multiprime for genome engineering by CRISPR/Cas9. |
| Databáze: | OpenAIRE |
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