Gene Expression Inside Liposomes: From Early Studies to Current Protocols.

Synthesizing proteins inside liposomes and other microcompartments is a well‐established practice. However, the origin of this research is not from the distant past, dating back to 1999–2004, when the first successful attempts were published. Protein synthesis inside artificial compartments is now under strong expansion in the context of synthetic biology (in bottom‐up approaches), and, in particular, it strongly contributes to the construction of artificial cell‐like systems. These systems, often called "synthetic cells", can be used to model cellular processes, including membrane‐centered ones. They are very innovative models that complement traditional studies and promise future applications. This review does not discuss all current directions in synthetic cell research; in particular, it does not include all kinds of artificial compartments. Instead, it is uniquely dedicated to the analysis of historical and technical developments of protein synthesis inside liposomes, highlighting a selected list of open questions. One of the goals is to note the importance of mastering liposome technology together with cell‐free systems for the successful realization of this specific type of synthetic cell. With this aim, four currently employed protocols are compared and discussed, with a major emphasis on the droplet transfer method, which is attractive due to its simplicity and encapsulation efficiency. [ABSTRACT FROM AUTHOR]