Optimization and quantification of protein synthesis inside liposomes

Journal of Liposome Research
Maryam AmidiEnrico Mastrobattista

Abstract

Synthetic biology aims at reprogramming existing, or creating new, biological systems, with the ultimate aim to obtain artificial cells whose functions can be tailored. For the latter, encapsulation of complex biochemical reactions into cell-sized compartments, such as liposomes, is required. Recently, several groups have demonstrated that proteins of interest can be produced de novo within liposomes by entrapping cell-free protein-synthesis systems and DNA templates inside liposomes. Although detectable, intraliposomal protein synthesis was generally poor. Here, we have optimized intraliposomal cell-free protein synthesis by changing several variables, including lipid composition as well as liposome, pyrophosphatase, and T7 RNA polymerase concentration. Further, by using an activity-based assay, we have quantified the amount of full-length protein that was produced from DNA templates inside liposomes before and after optimization of aforementioned variables. Based on the model protein beta-galactosidase, it is demonstrated that liposomal protein synthesis can yield microgram quantities of protein (30-40 microg/mL liposomes).

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Citations

Dec 22, 2012·Nanomedicine·Enrico Mastrobattista
Sep 29, 2011·Systems and Synthetic Biology·Maryam AmidiEnrico Mastrobattista
Jan 17, 2012·Angewandte Chemie·Zohreh NourianChristophe Danelon
Jun 8, 2011·Biotechnology Journal·Pasquale Stano
Sep 15, 2015·Angewandte Chemie·Vivienne ChanChristian J Kastrup
Sep 4, 2012·Journal of Controlled Release : Official Journal of the Controlled Release Society·Maryam AmidiEnrico Mastrobattista
May 15, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Koji NishimuraTetsuya Yomo
May 10, 2013·ACS Synthetic Biology·Zohreh Nourian, Christophe Danelon
Apr 9, 2019·Annals of the New York Academy of Sciences·Norikazu Ichihashi
Nov 30, 2011·Nature Reviews. Genetics·Wilfried Weber, Martin Fussenegger
Dec 17, 2019·Biotechnology and Bioengineering·Mohammad H Ayoubi-JoshaghaniRana Jahanban-Esfahlan

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