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|Title:||Forward chemical genetic screening|
Forward chemical genetics
Tagged chemical library
|Source:||Choi, H.,Kim, J.-Y.,Chang, Y.T.,Nam, H.G. (2014). Forward chemical genetic screening. Methods in Molecular Biology 1062 : 393-404. ScholarBank@NUS Repository. https://doi.org/10.1007/978-1-62703-580-4-21|
|Abstract:||Chemical genetics utilizes small molecules to perturb biological processes. Unlike conventional genetics methods, which involve the alteration of genetic information mostly with lasting effects, chemical genetics allows temporary and reversible alterations of biological processes. Furthermore, it enables the alteration of biological processes in a dose-dependent manner, providing an advantage over conventional genetics. In the present chapter, the general procedures of forward chemical genetic screening are described. Forward chemical genetic screening can be performed in three steps. The first step involves the identification of small molecules that induce phenotypic or physiological changes in a biological system from a chemical library. In the second step, cellular targets that interact with the isolated chemical, which are mostly proteins, are identified. Although several methods can be applied in the second step, the most common one is affinity pull-down assay using a target protein that binds to the isolated compound. However, affinity pull-down of a target protein is a formidable barrier in forward chemical genetics. We introduced a tagged chemical library approach that significantly facilitates the identification of target proteins. The third step consists of the validation of the target protein, which should include the assessment of target specificity. This step is critical because small molecules often show pleiotropic effects due to low specificity. The specificity test may include a competition assay using cold competitors and a genetic study using mutants or transgenic lines modified for the cellular target. © 2014 Springer Science+Business Media New York.|
|Source Title:||Methods in Molecular Biology|
|Appears in Collections:||Staff Publications|
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