ACEMBL Tool-Kits for High-Throughput Multigene Delivery and Expression in Prokaryotic and Eukaryotic Hosts

Adv Exp Med Biol. 2016:896:27-42. doi: 10.1007/978-3-319-27216-0_3.

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.

Keywords: Automation; Gene delivery; High-throughput; Membrane proteins; Metabolic engineering; Protein complexes; Robotics; Structural proteomics; Synthetic biology.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Automation, Laboratory
  • Eukaryotic Cells / metabolism*
  • Gene Expression Regulation
  • Gene Transfer Techniques*
  • Genetic Vectors
  • High-Throughput Screening Assays*
  • Humans
  • Multiprotein Complexes
  • Plasmids / genetics
  • Plasmids / metabolism
  • Prokaryotic Cells / metabolism*
  • Protein Conformation
  • Protein Engineering / methods*
  • Protein Multimerization
  • Protein Subunits
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Structure-Activity Relationship

Substances

  • Multiprotein Complexes
  • Protein Subunits
  • Recombinant Proteins