The New Age of Cell-Free Biology

Abstract

The cell-free molecular synthesis of biochemical systems is a rapidly growing field of research. Advances in the Human Genome Project, DNA synthesis, and other technologies have allowed the in vitro construction of biochemical systems, termed cell-free biology, to emerge as an exciting domain of bioengineering. Cell-free biology ranges from the molecular to the cell-population scales, using an ever-expanding variety of experimental platforms and toolboxes. In this review, we discuss the ongoing efforts undertaken in the three major classes of cell-free biology methodologies, namely protein-based, nucleic acids–based, and cell-free transcription–translation systems, and provide our perspectives on the current challenges as well as the major goals in each of the subfields.

Facile Formation of Giant Elastin-like Polypeptide Vesicles as Synthetic Cells

“…A distinct advantage of an ELP-based compartment is that these peptides can be genetically encoded and thus we can leverage the power of cell-free protein expression 26 to synthesize ELPs in situ. However, we do not expect the expressed ELP to selfassemble into cell-sized vesicles.…”

Engineering spatiotemporal organization and dynamics in synthetic cells

“…Recently, a DNA‐based cortex was incorporated into giant vesicles, endowing them with significantly higher mechanical stability toward osmotic shock (Kurokawa et al, 2017). Additionally, the powerful toolbox of DNA origami promises to enable the creation of structures supporting the membrane or even shaping the compartment (Dong et al, 2017; Noireaux & Liu, 2020). The use of DNA as a structural material can extend our degree of control on the shape of synthetic cells and improve the flexibility in the range of stresses and stimuli they can withstand and respond to.…”

“…In this regard, synthetic cells (also known as artificial cells) represent the natural evolution of biology as a science. Currently, synthetic cells are defined as encapsulated biochemical systems that can recapitulate one or a few of the functions that a natural cell can perform (Noireaux & Liu, 2020; Xu, Hu, & Chen, 2016; Figure 1). The engineering of synthetic cells can have a variety of purposes.…”

Cell mimicry as a bottom‐up strategy for hierarchical engineering ofnature‐inspiredentities

“…Another popular approach to provide AC functionality is by utilizing cell‐free expression systems within cell‐sized compartments, as reviewed in detail elsewhere (Jeong et al, 2019; Lyu et al, 2020; Noireaux & Liu, 2020; Yue, Zhu, & Kai, 2019). Incorporating the machinery for protein transcription‐translation (TX‐TL) allows for the AC to contain information in a similar manner to the natural cells by utilizing nucleic acids (DNA and RNA) as information carrying molecules and serve as a means to program and develop functions in the AC.…”