Cell Free Expression in Proteinosomes Prepared from Native Protein-PNIPAAm Conjugates

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Mengfei Gao - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Dishi Wang - , Chair of Organic Chemistry of Polymers, Leibniz Institute of Polymer Research Dresden, TUD Dresden University of Technology (Author)
  • Michaela Wilsch-Bräuninger - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Weihua Leng - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Jonathan Schulte - , Goethe University Frankfurt a.M. (Author)
  • Nina Morgner - , Goethe University Frankfurt a.M. (Author)
  • Dietmar Appelhans - , Leibniz Institute of Polymer Research Dresden, TUD Dresden University of Technology (Author)
  • T. Y.Dora Tang - , Max Planck Institute of Molecular Cell Biology and Genetics, Saarland University (Author)

Abstract

Towards the goal of building synthetic cells from the bottom-up, the establishment of micrometer-sized compartments that contain and support cell free transcription and translation that couple cellular structure to function is of critical importance. Proteinosomes, formed from crosslinked cationized protein-polymer conjugates offer a promising solution to membrane-bound compartmentalization with an open, semi-permeable membrane. Critically, to date, there has been no demonstration of cell free transcription and translation within water-in-water proteinosomes. Herein, a novel approach to generate proteinosomes that can support cell free transcription and translation is presented. This approach generates proteinosomes directly from native protein-polymer (BSA-PNIPAAm) conjugates. These native proteinosomes offer an excellent alternative as a synthetic cell chassis to other membrane bound compartments. Significantly, the native proteinosomes are stable under high salt conditions that enables the ability to support cell free transcription and translation and offer enhanced protein expression compared to proteinosomes prepared from traditional methodologies. Furthermore, the integration of native proteinosomes into higher order synthetic cellular architectures with membrane free compartments such as liposomes is demonstrated. The integration of bioinspired architectural elements with the central dogma is an essential building block for realizing minimal synthetic cells and is key for exploiting artificial cells in real-world applications.

Details

Original languageEnglish
Article number2300464
JournalMacromolecular bioscience
Volume24
Issue number3
Publication statusPublished - Mar 2024
Peer-reviewedYes

External IDs

PubMed 37925629

Keywords

Keywords

  • cell free expression, compartmentalisation, proteinosomes, synthetic cells