This book reviews cell-free production systems, exploring the frontiers in cellular engineering and biotechnology. With contributions from experts in the field, the book offers a comprehensive and up-to-date account of the latest advancements and practical applications. The volume covers a diverse range of topics, beginning with an in-depth analysis of cell-free display techniques for protein evolution, shedding light on the methodologies used to engineer proteins for diverse purposes, followed by an examination of bottom-up synthetic biology employing cell-free protein synthesis. Additionally, it investigates the intricacies of the cell-free synthesis of metalloproteins, elucidating the unique properties and functionalities of these biologically important molecules. In this book, particular attention is given to the integration of cell-free production systems with droplet microfluidics, a pioneering approach that has revolutionized research activities in both academic and industrial settings. Readers will also discover the latest advancements in cell-free protein synthesis and immobilization, and find out more about the eCell technology, which combines cell-free protein synthesis with bio-sensing and remediation, revolutionizing critical areas of study in biotechnology. Together with the companion volume entitled “Cell-free Production: System Development”, both books highlight the research progresses on the basic and applied research of cell-free production systems in the last few years, and are invaluable resources for scholars, researchers, and bioengineers. This book also appeals to enthusiasts of synthetic biology.