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

Cell Syst. 2019 Apr 24;8(4):338-344.e8. doi: 10.1016/j.cels.2019.03.005. Epub 2019 Apr 3.

Abstract

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 (URA3AI, KanMXAI, HphMXAI, and NatMXAI), 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.

Keywords: Cre; Saccharomyces cerevisiae, barcode sequencing; combinatorial screening; fitness; iSeq; interaction screening; landing pad; loxP; yeast.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fungal Proteins / chemistry
  • Fungal Proteins / metabolism
  • Gene Library
  • Protein Interaction Mapping / methods*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae
  • Sequence Analysis, DNA / methods*
  • Software*

Substances

  • Fungal Proteins
  • Recombinant Proteins