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Chemical Screening Using Cell-Free Egg Extract. | LitMetric

Chemical Screening Using Cell-Free Egg Extract.

Cold Spring Harb Protoc

Department of Cell and Developmental Biology, Vanderbilt Ingram Cancer Center, Vanderbilt, University Medical Center, Nashville, Tennessee 37232

Published: August 2018

Most drug screening methods use purified proteins, cultured cells, and/or small model organisms such as , zebrafish, flies, or nematodes. These systems have proven successes in drug discovery, but they also have weaknesses. Although purified cellular components allow for identification of compounds with activity against specific targets, such systems lack the complex biological interactions present in cellular and organismal screens. In vivo systems overcome these weaknesses, but the lack of cellular permeability, efflux by cellular pumps, and/or toxicity can be major limitations. egg extract, a concentrated and biologically active cytosol, can potentially overcome these weaknesses. Drug interactions occur in a near-physiological milieu, thereby functioning in a "truer" endogenous manner than purified components. Also, egg extract is a cell-free system that lacks intact plasma membranes that could restrict drug access to potential targets. Finally, egg extract is readily manipulated at the protein level: Proteins are easily depleted or added to the system, an important feature for analyzing drug effects in disease states. Thus, egg extract offers an attractive media for screening drugs that merges strengths of both in vitro and in vivo systems.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368528PMC
http://dx.doi.org/10.1101/pdb.prot098277DOI Listing

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