SLAS 2019 AI in Process Automation Symposium

Large scale cell based phenotypic screens of RNAi libraries are well established but can be limited by incomplete and unknown levels of gene knockdown. Here we present a platform for high content screening of an arrayed lentiviral CRISPR library for the whole human and mouse genomes which has recently been established by us. The use of automation allowed us to minimize handling and any potential exposure to the viral material as well as maximize the assay throughput. Here we present the methods used and example data from the complete process.

The automated steps include culturing, inoculation, staining and imaging, all in 1536 well plates. All liquid handling is performed within an enclosed JANUS workstation with attached carrousel and incubator. Imaging is performed with an opera phenix. Assay volumes have been trialled in the range of 5 - 8 ul per well, evaporation is largely mitigated by use of a filled moat in the plate skirting. Media exchange / washing is less prone to damaging the cell layer as result of such small assay volumes.

The data presented here is representative of the platforms use in whole genome phenotypic screening experiments. This platform represents a new stage of genome screening allowing rapid identification of gene targets for novel phenotypes. By interrogating each gene individually we can discern which target or pathway is involved in producing the phenotype of interest.