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

Journal Article · 03 April 2019 · Cell Systems

DOI: https://doi.org/10.1016/j.cels.2019.03.005 · OSTI ID:1529258

Liu, Xianan ^[1]; Liu, Zhimin ^[1]; Dziulko, Adam K. ^[1]; Li, Fangfei ^[1]; Miller, Darach ^[2]; Morabito, Robert D. ^[1]; Francois, Danielle ^[1]; Levy, Sasha F. ^[3]

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1. Stony Brook Univ., Stony Brook, NY (United States)
2. SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., Stanford, CA (United States)
3. Stony Brook Univ., Stony Brook, NY (United States); Joint Initiative for Metrology in Biology, Stanford, CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States); Stanford Univ., Stanford, CA (United States)

Here, 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_AI, KanMX_AI, HphMX_AI, and NatMX_AI), 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.

• Research Organization: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
• Sponsoring Organization: USDOE
• Grant/Contract Number: AC02-76SF00515; R01HG008354; U01HL127522
• OSTI ID: 1529258
• Journal Information: Cell Systems, Vol. 8, Issue 4; ISSN 2405-4712
• Publisher: ElsevierCopyright Statement
• Country of Publication: United States
• Language: English