Reconstituting Natural Cell Elements in Synthetic Cells

Abstract

Building a live cell from non‐living building blocks would be a fundamental breakthrough in biological sciences, and it would enable engineering new lineages of life, not directly descendant of the Last Universal Common Ancestor. Fully engineered synthetic cells will have architectures that can be radically different from the natural cells, yet most life processes reconstituted in synthetic cells so far are built from natural and biosimilar building blocks. Most natural processes have already been reconstituted in synthetic cell chassis. This paper summarizes recent advancements in using non‐living building blocks to reconstitute some of the most crucial features of living systems in a fully engineerable chassis of a synthetic cell.

T7Max transcription system

“…Cell free expression platforms find increasingly versatile applications in many areas of bioengineering, synthetic biology, and metabolic engineering [ 39 – 41 ]. Additionally, the focus for engineering synthetic minimal cells is on reconstituting in vitro translation reactions , most often with the use of a bacterial translation system and T7 RNA polymerase [ 42 , 43 ]. Here we demonstrated a simple technique to enable a significant increase in translation yield via a change of the T7 promoter sequence.…”

T7Max transcription system

“…[32][33][34] Additionally, the focus for engineering synthetic minimal cells is on reconstituting in vitro translation reactions , most often with the use of a bacterial translation system and T7 RNA polymerase. [35,36] Here we demonstrated a simple technique to enable a significant increase in translation yield via a change of the T7 promoter sequence.…”

A sustainable chemiosmotic strategy for driving solute transport in synthetic cells

“…This is an essential feature of living cells, not only to transport molecules in but also to pump metabolic end products out. Additionally, the components of the system presented here are proteins that can be expressed and regenerated from their genetic units by an encapsulated transcription-translation machinery, contributing to the construction of an autonomous synthetic cell (47,48). Besides the sustained chemiosmotic transport of nutrients, our study offers kinetic and mechanistic insights for the two protein components of the L-malate decarboxylation pathway (Figure 1 & 2).…”