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Programmable On-Chip Artificial Cell Producing Post-Translationally Modified Ubiquitinated Protein.

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Zilberzwige-Tal, Shai 
Toprakcioglu, Zenon  ORCID logo  https://orcid.org/0000-0003-1964-8432
Knowles, Tuomas PJ 
Gazit, Ehud 

Abstract

In nature, intracellular microcompartments have evolved to allow the simultaneous execution of tightly regulated complex processes within a controlled environment. This architecture serves as the blueprint for the construction of a wide array of artificial cells. However, such systems are inadequate in their ability to confine and sequentially control multiple central dogma activities (transcription, translation, and post-translational modifications) resulting in a limited production of complex biomolecules. Here, an artificial cell-on-a-chip comprising hierarchical compartments allowing the processing and transport of products from transcription, translation, and post-translational modifications through connecting channels is designed and fabricated. This platform generates a tightly controlled system, yielding directly a purified modified protein, with the potential to produce proteoform of choice. Using this platform, the full ubiquitinated form of the Parkinson's disease-associated α-synuclein is generated starting from DNA, in a single device. By bringing together all central dogma activities in a single controllable platform, this approach will open up new possibilities for the synthesis of complex targets, will allow to decipher diverse molecular mechanisms in health and disease and to engineer protein-based materials and pharmaceutical agents.

Description

Keywords

aptamers, cell free, microfluidics, post-translation modifications, synthetic biology

Journal Title

Small

Conference Name

Journal ISSN

1613-6810
1613-6829

Volume Title

Publisher

Wiley

Rights

All rights reserved
Sponsorship
This study was supported by the Adelis Foundation. S.Z.T. was supported by the Daniel Turnberg Travel Fellowship.