Title : Cellular Target Deconvolution of Small Molecules Using a Selection-Based Genetic Screening Platform - Zhao_2022_ACS.Cent.Sci_8_1424 |
Author(s) : Zhao J , Tang Z , Selvaraju M , Johnson KA , Douglas JT , Gao PF , Petrassi HM , Wang MZ , Wang J |
Ref : ACS Cent Sci , 8 :1424 , 2022 |
Abstract :
Small-molecule drug target identification is an essential and often rate-limiting step in phenotypic drug discovery and remains a major challenge. Here, we report a novel platform for target identification of activators of signaling pathways by leveraging the power of a clustered regularly interspaced short palindromic repeats (CRISPR) knockout library. This platform links the expression of a suicide gene to the small-molecule-activated signaling pathway to create a selection system. With this system, loss-of-function screening using a CRISPR single-guide (sg) RNA library positively enriches cells in which the target has been knocked out. The identities of the drug targets and other essential genes required for the activity of small molecules of interest are then uncovered by sequencing. We tested this platform on BDW568, a newly discovered type-I interferon signaling activator, and identified stimulator of interferon genes (STING) as its target and carboxylesterase 1 (CES1) to be a key metabolizing enzyme required to activate BDW568 for target engagement. The platform we present here can be a general method applicable for target identification for a wide range of small molecules that activate different signaling pathways. |
PubMedSearch : Zhao_2022_ACS.Cent.Sci_8_1424 |
PubMedID: 36313155 |
Zhao J, Tang Z, Selvaraju M, Johnson KA, Douglas JT, Gao PF, Petrassi HM, Wang MZ, Wang J (2022)
Cellular Target Deconvolution of Small Molecules Using a Selection-Based Genetic Screening Platform
ACS Cent Sci
8 :1424
Zhao J, Tang Z, Selvaraju M, Johnson KA, Douglas JT, Gao PF, Petrassi HM, Wang MZ, Wang J (2022)
ACS Cent Sci
8 :1424