Cell-free protein synthesis and assembly on a biochip

Yael Heyman, Amnon Buxboim, Sharon G. Wolf, Shirley S. Daube, Roy H. Bar-Ziv*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

46 Scopus citations

Abstract

Biologically active complexes such as ribosomes and bacteriophages are formed through the self-assembly of proteins and nucleic acids. Recapitulating these biological self-assembly processes in a cell-free environment offers a way to develop synthetic biodevices. To visualize and understand the assembly process, a platform is required that enables simultaneous synthesis, assembly and imaging at the nanoscale. Here, we show that a silicon dioxide grid, used to support samples in transmission electron microscopy, can be modified into a biochip to combine in situ protein synthesis, assembly and imaging. Light is used to pattern the biochip surface with genes that encode specific proteins, and antibody traps that bind and assemble the nascent proteins. Using transmission electron microscopy imaging we show that protein nanotubes synthesized on the biochip surface in the presence of antibody traps efficiently assembled on these traps, but pre-assembled nanotubes were not effectively captured. Moreover, synthesis of green fluorescent protein from its immobilized gene generated a gradient of captured proteins decreasing in concentration away from the gene source. This biochip could be used to create spatial patterns of proteins assembled on surfaces.

Original languageAmerican English
Pages (from-to)374-378
Number of pages5
JournalNature Nanotechnology
Volume7
Issue number6
DOIs
StatePublished - Jun 2012
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported by the Israel Science Foundation (Converging Technology programme) and the Minerva Foundation. The electron microscopy studies were conducted at the Irving and Cherna Moskowitz Center for Nano and Bio-Nano Imaging at the Weizmann Institute of Science. The authors thank T. Arad and V. Shinder for TEM assistance.

Fingerprint

Dive into the research topics of 'Cell-free protein synthesis and assembly on a biochip'. Together they form a unique fingerprint.

Cite this