生物
清脆的
功能基因组学
基因组学
比较基因组学
CRISPR干扰
基因组
计算生物学
细菌人工染色体
基因组编辑
基因
遗传学
作者
Henry H. Lee,Nili Ostrov,Brandon G. Wong,Michaela A. Gold,Ahmad S. Khalil,George M. Church
出处
期刊:Nature microbiology
日期:2019-04-08
卷期号:4 (7): 1105-1113
被引量:170
标识
DOI:10.1038/s41564-019-0423-8
摘要
The fast-growing Gram-negative bacterium Vibrio natriegens is an attractive microbial system for molecular biology and biotechnology due to its remarkably short generation time1,2 and metabolic prowess3,4. However, efforts to uncover and utilize the mechanisms underlying its rapid growth are hampered by the scarcity of functional genomic data. Here, we develop a pooled genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) screen to identify a minimal set of genes required for rapid wild-type growth. Targeting 4,565 (99.7%) of predicted protein-coding genes, our screen uncovered core genes comprising putative essential and growth-supporting genes that are enriched for respiratory pathways. We found that 96% of core genes were located on the larger chromosome 1, with growth-neutral duplicates of core genes located primarily on chromosome 2. Our screen also refines metabolic pathway annotations by distinguishing functional biosynthetic enzymes from those predicted on the basis of comparative genomics. Taken together, this work provides a broadly applicable platform for high-throughput functional genomics to accelerate biological studies and engineering of V. natriegens.
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