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.
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.
Cell Syst. 2019 Apr 24;8(4):338-344.e8. doi: 10.1016/j.cels.2019.03.005. Epub 2019 Apr 3.
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.
我们开发了一种用于酿酒酵母组合筛选的灵活工具包，该工具包可生成大型细胞文库，每个文库都经过独特的条形码标记，以标记 DNA 元件的组合。这个交互测序平台（iSeq 2.0）包括基因组着陆垫，通过质粒的顺序整合或酵母交配来组装组合，包含 15 个条形码质粒文库，其中包含分离的可选择标记（URA3、KanMX、HphMX 和 NatMX），以及大约 24000 个“双条形码”菌株的阵列，这些菌株可以使现有的酵母文库 iSeq 兼容。各种 DNA 元件与 iSeq 兼容：整合质粒上引入的 DNA、工程化的基因组修饰或整个遗传背景。DNA 元件文库是模块化和可互换的，任何两个文库都可以组合，使得 iSeq 能够通过短读测序执行许多新的组合筛选。
