Multi-functional genome-wide CRISPR system for high throughput genotype–phenotype mapping

Jiazhang Lian, Carl Schultz, Mingfeng Cao, Mohammad HamediRad and Huimin Zhao ()
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Jiazhang Lian: University of Illinois at Urbana-Champaign
Carl Schultz: University of Illinois at Urbana-Champaign
Mingfeng Cao: University of Illinois at Urbana-Champaign
Mohammad HamediRad: University of Illinois at Urbana-Champaign
Huimin Zhao: University of Illinois at Urbana-Champaign

Nature Communications, 2019, vol. 10, issue 1, 1-10

Abstract: Abstract Genome-scale engineering is an indispensable tool to understand genome functions due to our limited knowledge of cellular networks. Unfortunately, most existing methods for genome-wide genotype–phenotype mapping are limited to a single mode of genomic alteration, i.e. overexpression, repression, or deletion. Here we report a multi-functional genome-wide CRISPR (MAGIC) system to precisely control the expression level of defined genes to desired levels throughout the whole genome. By combining the tri-functional CRISPR system and array-synthesized oligo pools, MAGIC is used to create, to the best of our knowledge, one of the most comprehensive and diversified genomic libraries in yeast ever reported. The power of MAGIC is demonstrated by the identification of previously uncharacterized genetic determinants of complex phenotypes, particularly those having synergistic interactions when perturbed to different expression levels. MAGIC represents a powerful synthetic biology tool to investigate fundamental biological questions as well as engineer complex phenotypes for biotechnological applications.

Date: 2019
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DOI: 10.1038/s41467-019-13621-4

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