Bottom-up synthetic biology: reconstitution in space and time.

Bottom-up reconstituting well-characterized functional molecular entities, parts and modules towards a synthetic cell will give new insights into the mechanisms and origin of life. However, a remaining central challenge is how to organize cellular processes spatiotemporally from their component parts in vitro. Here, we review cutting edge tools and technologies that can facilitate such a bottom-up reconstitution towards a synthetic cell in space and time, particularly with regard to the following aspects: (1) reliable model membrane-environment and microenvironment; (2) dynamic genetic regulation and self-sustaining transcription and translation machinery; (3) spatially organized cytoskeleton that supports the biological architecture and cellular self-reproduction in 3D.