Since its recent discovery, CRISPR/Cas9 has triggered a revolution in biomedical research and is rapidly becoming an indispensible tool for many research applications. This system uses bacterially derived enzyme Cas9 and a gene-specific guide RNA molecule to introduce nicks in precisely defined genomic locations leading to a complete gene knockout. Although many researchers within and outside Australia are using CRISPR to achieve single gene knockouts in vitro and in vivo, utilising CRISPR for high-throughput screening promises a new era for uncovering novel biological relationships and decoding genome function. The first CRISPR unbiased screens displayed remarkable results with low levels of off-targets and biological variation, making the hit identification easier and potentially improving the success of their clinical translation. These screens, however, typically run in a pooled format, where all gene knockout constructs are assembled in a single large collection, and are interrogated in the context of gross overall cell survival by next-generation sequencing. Although valuable in drug discovery, such an approach severely limits the scope of biological questions asked and exploration of the basic mechanisms governing cellular processes. The Victorian Centre for Functional Genomics (VCFG) has established an arrayed CRISPR platform based on the synthetic CRISPR reagents, which enables individual gene knockout in a single well coupled with versatile downstream analyses using high-content microscopy. The synthetic CRISPRs induce highly efficient and rapid gene knockout, offering several advantages over vector-based systems: streamlined experimental setup, short assay length, strong phenotypic penetrance in the absence of selection, and short half-life of CRISPR constructs within the cell. In addition, drawing on our extensive experience with liquid handling robotics, we developed fully automated pipelines for the delivery of synthetic CRISPRs into the cells as well as all subsequent analyses, making it an ideal tool for both discovery and validation work.