Reconstituting Natural Cell Elements in Synthetic Cells

2021 ◽  
Vol 5 (3) ◽  
pp. 2000188 ◽  
Author(s):  
Nathaniel J. Gaut ◽  
Katarzyna P. Adamala
Keyword(s):  
Synthetic Cells ◽  
Natural Cell

scholarly journals Facile Formation of Giant Elastin-like Polypeptide Vesicles as Synthetic Cells

2021 ◽  
Author(s):  
Bineet Sharma ◽  
Yutao Ma ◽  
Andrew Ferguson ◽  
Allen Liu
Keyword(s):  
Host Cell ◽  
Lipid Vesicles ◽  
Free Expression ◽  
Polymer Conjugates ◽  
Protein Polymer ◽  
Synthetic Cell ◽  
Synthetic Cells ◽  
Transfer Method ◽  
Physical And Chemical ◽  
Natural Cell

Creating a suitable compartment for synthetic cells has led the exploration of different cell chassis materials from phospholipids to polymer to protein-polymer conjugates. Currently, the majority of cell-like compartments are made of lipid molecules as the resulting membrane resembles that of a natural cell. However, cell-sized lipid vesicles are prone to physical and chemical stresses and can be unstable in hosting biochemical reactions within. Recently, peptide vesicles that are more robust and stable were developed as a new chassis material for synthetic cells. Here we demonstrate the facile and robust generation of giant peptide vesicles made of elastin-like polypeptides (ELPs) by using an emulsion transfer method. We show that these peptide vesicles can stably encapsulate molecules and can host cell-free expression reactions. We also demonstrate membrane incorporation of another amphiphilic ELP into existing peptide vesicles. Since ELPs are genetically encoded, the approaches presented here provide exciting opportunities to engineer synthetic cell membranes.

scholarly journals Facile Formation of Giant Elastin-like Polypeptide Vesicles as Synthetic Cells

2021 ◽  
Author(s):  
Bineet Sharma ◽  
Yutao Ma ◽  
Andrew Ferguson ◽  
Allen Liu
Keyword(s):  
Host Cell ◽  
Lipid Vesicles ◽  
Free Expression ◽  
Polymer Conjugates ◽  
Protein Polymer ◽  
Synthetic Cell ◽  
Synthetic Cells ◽  
Transfer Method ◽  
Physical And Chemical ◽  
Natural Cell

Creating a suitable compartment for synthetic cells has led the exploration of different cell chassis materials from phospholipids to polymer to protein-polymer conjugates. Currently, the majority of cell-like compartments are made of lipid molecules as the resulting membrane resembles that of a natural cell. However, cell-sized lipid vesicles are prone to physical and chemical stresses and can be unstable in hosting biochemical reactions within. Recently, peptide vesicles that are more robust and stable were developed as a new chassis material for synthetic cells. Here we demonstrate the facile and robust generation of giant peptide vesicles made of elastin-like polypeptides (ELPs) by using an emulsion transfer method. We show that these peptide vesicles can stably encapsulate molecules and can host cell-free expression reactions. We also demonstrate membrane incorporation of another amphiphilic ELP into existing peptide vesicles. Since ELPs are genetically encoded, the approaches presented here provide exciting opportunities to engineer synthetic cell membranes.

scholarly journals Giant Vesicles Produced with Phosphatidylcholines (PCs) and Phosphatidylethanolamines (PEs) by Water-in-Oil Inverted Emulsions

Life ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 223
Author(s):  
Boying Xu ◽  
Jinquan Ding ◽  
Jian Xu ◽  
Tetsuya Yomo
Keyword(s):  
Encapsulation Efficiency ◽  
Analytical Study ◽  
Giant Vesicles ◽  
Imaging Flow Cytometry ◽  
Artificial Cell ◽  
Transfer Method ◽  
Long Term Preservation ◽  
Natural Cell ◽  
Cell Construction

(1) Background: giant vesicles (GVs) are widely employed as models for studying physicochemical properties of bio-membranes and artificial cell construction due to their similarities to natural cell membranes. Considering the critical roles of GVs, various methods have been developed to prepare them. Notably, the water-in-oil (w/o) inverted emulsion-transfer method is reported to be the most promising, owning to the relatively higher productivity and better encapsulation efficiency of biomolecules. Previously, we successfully established an improved approach to acquire detailed information of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)-derived GVs with imaging flow cytometry (IFC); (2) Methods: we prepared GVs with different lipid compositions, including phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), and PC/PE mixtures by w/o inverted emulsion methods. We comprehensively compared the yield, purity, size, and encapsulation efficiency of the resulting vesicles; (3) Results: the relatively higher productivities of GVs could be obtained from POPC, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE), DOPC: DLPE (7:3), and POPC: DLPE (6:4) pools. Furthermore, we also demonstrate that these GVs are stable during long term preservation in 4 °C. (4) Conclusions: our results will be useful for the analytical study of GVs and GV-based applications.

scholarly journals Heterogeneous multi-compartmental hydrogel particles as synthetic cells for incompatible tandem reactions

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Hongliang Tan ◽  
Song Guo ◽  
Ngoc-Duy Dinh ◽  
Rongcong Luo ◽  
Lin Jin ◽  
...  
Keyword(s):  
Tandem Reactions ◽  
Hydrogel Particles ◽  
Synthetic Cells

Synthetic Cells

JAMA ◽  
2010 ◽  
Vol 304 (2) ◽  
pp. 148
Author(s):  
Mike Mitka
Keyword(s):  
Synthetic Cells

scholarly journals General principles for the formation and proliferation of a wall-free (L-form) state in bacteria

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Romain Mercier ◽  
Yoshikazu Kawai ◽  
Jeff Errington
Keyword(s):  
Cell Wall ◽  
Molecular Biology ◽  
Gram Negative Bacteria ◽  
Systematic Analysis ◽  
Gram Negative ◽  
Membrane Blebbing ◽  
Precursor Synthesis ◽  
Peptidoglycan Cell Wall ◽  
Synthetic Cells ◽  
Opening Up

The peptidoglycan cell wall is a defining structural feature of the bacterial kingdom. Curiously, some bacteria have the ability to switch to a wall-free or ‘L-form’ state. Although known for decades, the general properties of L-forms are poorly understood, largely due to the lack of systematic analysis of L-forms in the molecular biology era. Here we show that inhibition of peptidoglycan precursor synthesis promotes the generation of L-forms from both Gram-positive and Gram-negative bacteria. We show that the L-forms generated have in common a mechanism of proliferation involving membrane blebbing and tubulation, which is dependent on an altered rate of membrane synthesis. Crucially, this mode of proliferation is independent of the essential FtsZ based division machinery. Our results suggest that the L-form mode of proliferation is conserved across the bacterial kingdom, reinforcing the idea that it could have been used in primitive cells, and opening up its use in the generation of synthetic cells.

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