iSeq 2.0: A Modular and Interchangeable Toolkit for Interaction Screening in Yeast.

Cell Syst

Department of Biochemistry, Stony Brook University, Stony Brook, NY 11794-5215, USA; Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY 11794-5252, USA; Department of Applied Mathematics and Statistics, Stony Brook University, Stony Brook, NY 11794-5215, USA; Joint Initiative for Metrology in Biology, Stanford, CA 94305-4245, USA; SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA; Department of Genetics, Stanford University, Stanford, CA 94305-5120, USA. Electronic address:

Published: April 2019

We developed a flexible toolkit for combinatorial screening in Saccharomyces cerevisiae, which generates large libraries of cells, each uniquely barcoded to mark a combination of DNA elements. This interaction sequencing platform (iSeq 2.0) includes genomic landing pads that assemble combinations through sequential integration of plasmids or yeast mating, 15 barcoded plasmid libraries containing split selectable markers (URA3, KanMX, HphMX, and NatMX), and an array of ∼24,000 "double-barcoder" strains that can make existing yeast libraries iSeq compatible. Various DNA elements are compatible with iSeq: DNA introduced on integrating plasmids, engineered genomic modifications, or entire genetic backgrounds. DNA element libraries are modular and interchangeable, and any two libraries can be combined, making iSeq capable of performing many new combinatorial screens by short-read sequencing.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6483859PMC
http://dx.doi.org/10.1016/j.cels.2019.03.005DOI Listing

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