Three-way combinatorial CRISPR screen for analysing interactions among druggable targets

We established CRISPR-based multi-gene knockout screening system and new toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. This system is able to identify not only pairwise but also three-way synergistic therapeutic target combinations and validate double and triple combination regimens for suppression on cancer cell growth and protection against Parkinson’s disease-associated toxicity. Moreover, it is applied for rapid screening of disease-alleviating gene knockouts to study high-order genetic interactions and uncovering the rare synergistic interactions between druggable targets.