title: ACEMBL Tool-Kits for High-Throughput Multigene Delivery and Expression in Prokaryotic and Eukaryotic Hosts.
abstract: Multicomponent biological systems perform a wide variety of functions and are crucially important for a broad range of critical health and disease states. A multitude of applications in contemporary molecular and synthetic biology rely on efficient, robust and flexible methods to assemble multicomponent DNA circuits as a prerequisite to recapitulate such biological systems in vitro and in vivo. Numerous functionalities need to be combined to allow for the controlled realization of information encoded in a defined DNA circuit. Much of biological function in cells is catalyzed by multiprotein machines typically made up of many subunits. Provision of these multiprotein complexes in the test-tube is a vital prerequisite to study their structure and function, to understand biology and to develop intervention strategies to correct malfunction in disease states. ACEMBL is a technology concept that specifically addresses the requirements of multicomponent DNA assembly into multigene constructs, for gene delivery and the production of multiprotein complexes in high-throughput. ACEMBL is applicable to prokaryotic and eukaryotic expression hosts, to accelerate basic and applied research and development. The ACEMBL concept, reagents, protocols and its potential are reviewed in this contribution.
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227, Dortmund, Germany.
authors.forename: Yan
authors.initials: Y
authors.lastname: Nie
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Chaillet
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Department of Biochemistry, Freie Universität Berlin, Takustrasse 6, 14195, Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Becke
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Novartis Institutes for BioMedical Research, Novartis Campus, 4056, Basel, Switzerland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Haffke
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.
authors.forename: Martin
authors.initials: M
authors.lastname: Pelosse
authors.affiliation: Geneva Biotech SARL, Avenue de la Roseraie 64, 1205, Genève, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Fitzgerald
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Collinson
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France.|School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK.
authors.forename: Christiane
authors.initials: C
authors.lastname: Schaffitzel
authors.affiliation: European Molecular Biology Laboratory, Grenoble Outstation, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France. iberger@embl.fr.|Unit of Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, UMI 3265, 71 avenue des Martyrs, 38042, Grenoble Cedex 9, France. iberger@embl.fr.|School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK. iberger@embl.fr.
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
mesh_terms: D000818:Animals; D057205:Automation, Laboratory; D005057:Eukaryotic Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D057166:High-Throughput Screening Assays; D006801:Humans; D046912:Multiprotein Complexes; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011387:Prokaryotic Cells / Q000378:metabolism*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D055503:Protein Multimerization; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D046912:Multiprotein Complexes; D021122:Protein Subunits; D011994:Recombinant Proteins
keywords: Automation; Gene delivery; High-throughput; Membrane proteins; Metabolic engineering; Protein complexes; Robotics; Structural proteomics; Synthetic biology