Cell free translation system for compound screening and related uses

Abstract

The invention provides a cell-free system comprising not more than about 5% wheat germ extract for expressing proteins such as viral proteins and proteins required for viral capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids, are provided. Further provided are methods for expressing proteins such as viral proteins, proteins required for capsid assembly, and proteins that assemble into multiprotein complexes in a manner analogous to viral capsids using a cell-free system comprising not more than about 5% wheat germ extract. Further provided are methods to assay for compounds that modulate viral protein, viral capsid assembly, and assembly of proteins into multiprotein complexes whose disruption can ameliorate bacterial, parasitic, metabolic, oncologic, immunologic, or CNS disease in a cell-free system comprising not more than about 5% wheat germ extract.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS[0001]The present application claims the benefit of U.S. Provisional Application No. 61 / 514,825, filed Aug. 3, 2011, the content of which is expressly incorporated herein by reference in its entirely for all purposes.STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT[0002]Not applicableFIELD OF THE INVENTION[0003]The present invention relates to a wheat germ extract for use in compositions for cell-free translation, methods of expressing proteins using a composition including wheat germ extract, methods of assembling viral capsids, and methods of detecting compounds that modulate protein-protein interactions including but not limited to the assembly of viral capsids.BACKGROUND OF THE INVENTION[0004]Cell-free translation systems utilize cellular extracts to express proteins of interest. In this paradigm, an in vitro transcribed or isolated RNA template is added to the cellular extract along with additio...

AI Technical Summary

Benefits of technology

[0077]An advantage of using the cell-free system of the present invention is that the process of capsid formation is slowed down, thus allowing for the targeting of capsid assembly processes for modulators of capsid assembly. An advantage of using cell-free systems of the invention, such as those having about 5% wheat germ extract, is an increased sensitivity for detecting compounds that otherwise would not be detected at higher wheat germ concentrations.

Problems solved by technology

Currently used cell-free systems do not show useful correlation between the ability of therapeutic agents that disrupt capsid assembly in vitro and the ability of these therapeutic agents to prevent virus replication.

Images