Zhou, P., Chan, B.K.C., Wan, Y.K., Yuen, C.T.L., Choi, G.C.G., Li, X., Tong, C.S.W., Zhong, S.S.W., Sun, J., Bao, Y., Mak, S.Y.L., Chow, M.Z.Y., Khaw, J.V., Leung, S.Y., Zheng, Z., Cheung, L.W.T., Tan, K., Wong, K.H., Chan, H.Y.E., Wong, A.S.L. (2020). A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets.  Cell Rep. 32(6): 108020.

FBrf0246426

Research paper

We present a CRISPR-based multi-gene knockout screening system and toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. We apply this system for systematically identifying not only pairwise but also three-way synergistic therapeutic target combinations and successfully validate double- and triple-combination regimens for suppression of cancer cell growth and protection against Parkinson's disease-associated toxicity. This system overcomes the practical challenges of experimenting on a large number of high-order genetic and drug combinations and can be applied to uncover the rare synergistic interactions between druggable targets.