Both our team (1) and others (2) have previously used large-scale CRISPR-Cas9 screens to prioritise novel therapeutic targets in cancer. Building on this, we have established a next-generation dual guide vector tool to systematically knock out tens of thousands of gene pairs and identify therapeutically relevant combinations in cancers of unmet clinical need. Our vector system is composed of two sgRNAs with a tRNA spacer, which allows processing by the cell’s endogenous tRNA processing system, enabling simultaneous perturbation of two targets. 

Here, we present ongoing collaborative efforts to validate synergies between potential target combination candidates in two separate, but complementary workflows. High throughput arrayed CRISPR screening, based on systematic target prioritisation takes place in parallel to a lower throughput approach, using a more target-focused and biomarker-driven experimentation. Both approaches utilise orthogonal methodologies to validate target combinations emerging from pooled CRISPR screens, and individually offer their own advantages, as well as generating additional supporting evidence for targets nominated for drug discovery.