DNA Lattice-Encoded Information for Genotype-to-Phenotype Evolution of Self-Replicating Synthetic Cells

Informazioni relative al progetto

DNA-ENC SYNCELLS

ID dell’accordo di sovvenzione: 792270

DOI

10.3030/792270

Progetto concluso il 31 Ottobre 2019

Data della firma CE 28 Febbraio 2018

Data di avvio 1 Marzo 2018

Data di completamento 29 Febbraio 2020

Finanziato da

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Costo totale

€ 159 460,80

Contributo UE

€ 159 460,80

Coordinato da

MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV
Germany

Periodic Reporting for period 1 - DNA-ENC SYNCELLS (DNA Lattice-Encoded Information for Genotype-to-Phenotype Evolution of Self-Replicating Synthetic Cells)

Periodo di rendicontazione: 2018-03-01 al 2020-02-29

Sintesi del contesto e degli obiettivi generali del progetto

Living cells may be the most sophisticated example of functional machines operating on the nano- and microscale. But is it possible engineer synthetic analogues in the laboratory? In this fellowship, we propose a strategy to engineer cells de novo, by merging two precision technologies, microfluidics and DNA nanotechnology, to position and manipulate functional parts in space and time [1]. In particular, we use DNA as a near-universal linker to functionalize microfluidic compartments. Using this DNA handle approach, we demonstrate the stimuli-responsive attachment of natural and synthetic subcellular components. We further employ DNA to construct functional parts, including ion channels or a pH-responsive DNA-based cytoskeleton mimic, which serves as a stabilizing cortex. Following passive encapsulation, we actuate DNA nanostructures in microfluidic or lipid-based compartments to assemble dynamic systems with structural reconfigurability. By the integration of plasmonic probes we achieve real-time optical feedback to monitor the dynamics upon external stimulation. These unique tools bridging the mico- and nanoscale will enable synthetic cellular systems, which are not merely copies of nature’s own example of life, but prescribe a direction towards synthetic cellular machines, ultimately applicable as active probes inside the human body.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

First, I established a new method to form giant unilamellar vesicles as cell-like compartments in a simple one-pot reaction. This technique will directly benefit laboratories around the world (ACS Synth. Biol.). Second, I invented a near-universal method to functionalize microfluidic droplets via DNA handles – now filed to the European Patent Office. Having guided my Master student through the attachment of various functional groups and components, I am the co-corresponding author on the manuscript in Adv. Funct. Mater. I then employed proteorhodopsin-expressing bacteria in synthetic cells to trigger the reversible assembly and disassembly of a pH-responsive DNA-based cytoskeleton upon illumination. For the first time, bacteria are used as organelles in synthetic cells (to be submitted). Remarkably, I then demonstrated autonomous division of synthetic cells. I further established a small group with independent collaborations and laid out my view on engineering synthetic cells in a review featured on the cover of Trends in Biotechnology.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

We combined DNA nanotechnology and droplet-based microfluidics. Thereby, we established a new direction in synthetic biology, paving the way towards truely synthetic cells assembled de novo. Tangible applications were realized with a new technique to functionalize microfluidic droplets (patent pending).

Functionalization of microfluidic droplets with DNA