title: Light it up: highly efficient multigene delivery in mammalian cells.
abstract: Multigene delivery and expression systems are emerging as key technologies for many applications in contemporary biology. We have developed new methods for multigene delivery and expression in eukaryotic hosts for a variety of applications, including production of protein complexes for structural biology and drug development, provision of multicomponent protein biologics, and cell-based assays. We implemented tandem recombineering to facilitate rapid generation of multicomponent gene expression constructs for efficient transformation of mammalian cells, resulting in homogenous cell populations. Analysis of multiple parameters in living cells may require co-expression of fluorescently tagged sensors simultaneously in a single cell, at defined and ideally controlled ratios. Our method enables such applications by overcoming currently limiting challenges. Here, we review recent multigene delivery and expression strategies and their exploitation in mammalian cells. We discuss applications in drug discovery assays, interaction studies, and biologics production, which may benefit in the future from our novel approach.
authors: European Molecular Biology Laboratory-EMBL, Grenoble, France.
authors: Simon
authors: S
authors: Trowitzsch
authors: Martin
authors: M
authors: Klumpp
authors: Ralf
authors: R
authors: Thoma
authors: Jean-Philippe
authors: JP
authors: Carralot
authors: Imre
authors: I
authors: Berger
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008322:Mammals; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D018079:Receptors, GABA / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; D018079:Receptors, GABA