Functionalizing cell-mimetic giant vesicles with encapsulated bacterial biosensors.

Tatiana Trantidou,Karen Polizzi,Yuval Elani,Oscar Ces,Linda Dekker

doi:10.1098/rsfs.2018.0024

Abstract

The design of vesicle microsystems as artificial cells (bottom-up synthetic biology) has traditionally relied on the incorporation of molecular components to impart functionality. These cell mimics have reduced capabilities compared with their engineered biological counterparts (top-down synthetic biology), as they lack the powerful metabolic and regulatory pathways associated with living systems. There is increasing scope for using whole intact cellular components as functional modules within artificial cells, as a route to increase the capabilities of artificial cells. In this feasibility study, we design and embed genetically engineered microbes (Escherichia coli) in a vesicle-based cell mimic and use them as biosensing modules for real-time monitoring of lactate in the external environment. Using this conceptual framework, the functionality of other microbial devices can be conferred into vesicle microsystems in the future, bridging the gap between bottom-up and top-down synthetic biology.

Highlights

In recent years, there has been an upsurge in interest surrounding the design and construction of functional vesicle-based systems
License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited. We demonstrate this potential by designing genetically engineered microbes and using them as biosensing modules that are encapsulated within a vesicle chassis
Strains were maintained on Luria– Bertani (LB) agar containing 100 mg ml21 ampicillin and grown aerobically for 16 h at 378C

Summary

Introduction

There has been an upsurge in interest surrounding the design and construction of functional vesicle-based systems. The components responsible for their functionality are more diverse These include: ligands that decorate the membrane (e.g. DNA sticky ends, antibodies, tethered small molecules, nanoparticles, polymerizable amphiphiles) [7,12,13,14,15]; membrane-spanning structures (e.g. natural and engineered protein pores, pumps and channels, DNA origami) [16 –18]; and biologically derived material encapsulated in the vesicle lumen (e.g. enzymes, cytoskeleton components, polymerases and coupled transcription/translation machinery) [6,19,20,21,22]. This varied repertoire of building blocks has allowed investigators to engineer vesicles capable of mimicking an assortment of cellular behaviours [23], and of successfully interfacing cell and vesicle communities together [24]

Methods

Results

Conclusion