Multi-functional genome-wide CRISPR system for high throughput genotype–phenotype mapping
| Autor: | Mohammad HamediRad, Mingfeng Cao, Huimin Zhao, Jiazhang Lian, J. Carl Schultz |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine CRISPR-Cas systems Genotype Science General Physics and Astronomy Saccharomyces cerevisiae Computational biology Biology 01 natural sciences Genome Article General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Synthetic biology Gene Expression Regulation Fungal 010608 biotechnology CRISPR Genomic library lcsh:Science Gene Gene Editing Genomic Library Multidisciplinary Fungal genetics Magic (programming) Chromosome Mapping High-Throughput Nucleotide Sequencing Genomics General Chemistry Phenotype High-Throughput Screening Assays 030104 developmental biology lcsh:Q Genome Fungal Metabolic engineering Biotechnology |
| Zdroj: | Nature Communications, Vol 10, Iss 1, Pp 1-10 (2019) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | 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. Genome-scale engineering is generally limited to single methods of alteration such as overexpression, repression or deletion. Here the authors present a tri-functional CRISPR system that can engineer complex synergistic interactions in a genome-wide manner. |
| Databáze: | OpenAIRE |
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