Double Plasmid Landing Pad

Method of creating a double barcode library in order to evaluate genetic or protein interactions on a large scale in cell pools        Background: The recent innovation of molecular barcodes has increased High-Throughput Combinatorial Screening (HTCS) efficiency by allowing the simultaneous measurement of many combinations in pools of cells. Barcoded cells can be grown together and relative barcode abundances can be measured over time by amplicon sequencing, allowing fitness calculations at a cost that is orders of magnitude lower than one-at-a-time methods. However, large-scale bar-seq approaches are not widely used because a unique barcode must be constructed for each DNA element or combination, they do not function with the yeast mating method, and the entire library must be constructed again for every interaction in a new environment or cell type. Technology Overview: Dr. Sasha Levy, Assistant Professor at Stony Brook University developed a method that allows one to efficiently construct, in any cell type, >10 million combinations of DNA elements, each combination of which is uniquely identified by a double barcode that resides in the genome. Large double barcode pools can subsequently be used with Barcode-seq approaches to simultaneously assay each combination cheaply using next generation sequencing. Because this technology acts as an interaction sequencing platform, it will have broad applications in basic research and industry including high-throughput protein or genetic interaction screens, double gene deletion screens, drug screens, and candidate gene identification screens. Advantages: Not yeast specific Higher throughput construction of interaction pairs Faster Cheaper Higher Efficiency Many uses Can be used repeatedly at low cost Applications: Research Tool, drug discovery Intellectual Property Summary: provisional application Stage of Development: Research Licensing Potential: Licensing Status: Available for License Additional Information: Protein-protein interaction, Genetic interaction, Drug screening, High throughput, Barcode Sequencing https://stonybrook.technologypublisher.com/files/sites/research-tools-and-reagents1.jpg , https://stonybrook.technologypublisher.com/files/sites/screens-and-assays1.jpg
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