Drug Discovery
Multiparametric screening can be a powerful
application of arrayed screening, and since the pooled approach is less compatible with multi- plexed readouts, this can be a valuable tool for researchers seeking complex phenotypic data from their screen. For instance, one study evaluated the impact of gene knock-out on cell cycle progression through the use of synthetic CRISPRko reagents. The researchers used a high-content microscopy readout to classify cell cycle state coupled to a novel analytical process to assimilate parameters for robust hit ID29. The arrayed screening approach can even be extended to evaluate cell responses to gene modulations in difficult-to-edit model systems, such as primary T cells30. This highly impressive approach not only demonstrates the value of arrayed screening platform data, but also brings the technology to T-cells which have proven challenging to interrogate by established CRISPR screening techniques. This cell type is proving crucial for drug discovery in immuno- oncology, and arrayed screening provides the opportunity to explore T-cell impact in co-culture systems and evaluate non-cell autonomous effects with robust genetic perturbation technologies. Overall, arrayed CRISPR screening tools allow
researchers to conduct a very diverse range of com- plex assays and readouts including measuring changes in biomarker expression, localisation or cell and organelle morphology. The possibility of highly-multiplexed assay readouts also significant- ly increases the value of arrayed approaches.
Conclusions CRISPR has undoubtedly provided researchers with augmented tools for functional genomics, and with the continued implementation of new adapta- tions and innovations it is an exciting time to be a gene editing scientist. The coupling of phenotypic analysis to CRISPR-based screening has added promising value to the applications of these tech- nologies to more complex disease areas. In an age of personalised medicine, functional genomics is key and creativity, expertise and pragmatism in screening are crucial if these tools are to yield ben- efits for patient outcomes.
DDW
Dr Steffen Lawo is a Team Leader in Functional Genomic Screening at Horizon Discovery. At Horizon, Steffen led the development and imple- mentation of pooled CRISPRko screening and the establishment of this platform at Horizon. He has specific expertise in pooled and arrayed CRISPR and RNAi screening campaigns with complex,
Drug Discovery World Fall 2018 57
high-content phenotypic readouts. Before joining Horizon, Steffen earned his PhD in Molecular Genetics at the University of Toronto.
Dr Carlos le Sage is a Team Leader in Functional Genomic Screening at Horizon Discovery. At Horizon, Carlos focuses on discovering the mecha- nism of action of newly-developed drugs and defin- ing new synthetic lethal interactions in defined mutant-driven cancer genomes, through unbiased genetic perturbation screens. Prior to joining Horizon Discovery, Carlos completed postdoctoral research with Steve Jackson at the Gurdon Institute in Cambridge. He holds a PhD in Biology from the Netherlands Cancer Institute through Leiden University.
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