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High thoughput molecular function assays on the frontier of big data

Project: Tec Talk

Topics: Bioinformatics

Investigation of biological systems has historically focused on one-off types of experiments. The advent of more complex instrumentation and computer-based analyses has stimulated the development of impactful high-throughput techniques over the past few decades. A few examples of the major techniques are DNA sequencing, RNA sequencing, genome wide RNAi screens, and high content screens. These techniques enable the omics era of science with widespread analysis of genomes, proteomes, metabolomes, etc. Although there are now about a dozen major high throughput technologies in routine use, There is no high-throughput technology to assay different types of molecular functions with libraries of macromolecules in mammalian cells. This gap is critical because the molecular functions are the most essential to understand mechanisms, the etiology of disease, and for developing therapeutics. Our laboratory has developed an innovative pipeline combining gene editing, fluorescent reporters, flow cytometry, cell sorting, barcoding, and next-generation sequencing (NGS) into a New technology we call the GigaAssay. The GigaAssay can concurrently assay 100,000s of mammalian cells and replicates for effects of a variable in a library upon molecular functions. This single cell approach does not necessitate separating cells as done in a droplet generator. In our proof-of-principle example, we measured the transcriptional activity of ~60,000 Tat missense variants in one experiment with the GigaAssay; Tat is the HIV transcription factor. This technology will vastly accelerate the rate of study of cellular and molecular biology and the etiology of disease.

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