Cell-Free Production Platforms for Developing Tools and Applications in Pharmacology

Cell-free protein synthesis (CFPS) represents a rapidly growing multifaceted platform that is highly complementary to conventional cell-based biomolecule production technologies. CFPS systems are open and accessible throughout the reaction and tolerate a wide variety of supplements and modifications including small molecules, catalytic enzymes, redox tuning or artificial hydrophobic environments. These unique characteristics allow one to individually adjust reaction conditions according to specific requirements of a target, in view of production efficiency, folding, stability, or function. Furthermore, the complete control over reaction components enables efficient and cost-effective strategies for the labelling and modification of synthesized proteins. CFPS can be performed in diverse modes and scales from nanoliter reactions in microtiter formats up to multi-liter volumes in fermenters. The largely reduced metabolic activity in CFPS eliminates formerly significant bottlenecks in protein production such as DNA template instability, background expression and induction problems, as well as inefficient transport and metabolic conversion of supplied additives. CFPS has therefore evolved as a primary choice for the production of membrane proteins, toxins and complexes, for the labelling of proteins, and for the generation of artificial protein assemblies. In combination with its competitive speediness and high reproducibility, CFPS represents the most versatile biomolecule production and modification system currently available. It is therefore not surprising that CFPS is expected to have a highly innovative potential to impact biopharmaceutical research. New approaches and applications such as nucleic acid circuits, protein evolution, nanomachine design, and synthesis of virus-like particles for vaccination are continuously documented and indicate a bright future of CFPS.

This Research Topic intends to focus on articles and reviews having pharmacological relevance and covering case studies of applications, development of new approaches and strategies, as well as implementing different CFPS systems based on prokaryotic or eukaryotic cell lysates. The Topic will highlight the current and emerging synergies obtained by combining CFPS systems, nanotechnologies and pharmacological research. Submissions suitable for consideration may include: (i) Synthesis of assemblies, proteins or small molecules for structural or functional studies; (ii) Design of modified or new biosynthetic circuits; (iii) Synthesis of biomolecules in artificial environments; (iv) Development and optimization of cell-free production systems; (v) Throughput screening, array-technologies or preparative scale applications; (vi) Metabolic engineering in cell-free lysates; (vii) Protein evolution and modulation of protein folding and function; (viii) Protein labelling and expanding the genetic code; (ix) Production of difficult targets such as membrane proteins, toxins or peptides; (x) Artificial cells; (xi) Engineering post-translational modifications of targets.