Effects of CRISPR/Cas9 targeting of the myo-inositol biosynthesis pathway on hyper-osmotic tolerance of tilapia cells

被引：2
|
作者：
Hamar, Jens [1 ,2 ]
Cnaani, Avner [3 ]
Kultz, Dietmar [1 ,2 ]
机构：
[1] Univ Calif Davis, Dept Anim Sci, Meyer Hall,1 Shields Ave, Davis, CA 95616 USA
[2] Univ Calif Davis, Genome Ctr, Meyer Hall,1 Shields Ave, Davis, CA 95616 USA
[3] Agr Res Org, Inst Anim Sci, Volcani Ctr, Dept Poultry & Aquaculture, POB 15159, IL-7528809 Rishon Leziyyon, Israel
来源：
GENOMICS | 2024年 / 116卷 / 03期
基金：
美国国家科学基金会;
关键词：
CRISPR/Cas9; Osmoregulation; Salinity; Myo-inositol; Oreochromis tilapia; PENTOSE-PHOSPHATE PATHWAY; NILE TILAPIA; OREOCHROMIS-NILOTICUS; ENVIRONMENTAL SALINITY; VOLUME REGULATION; MYOINOSITOL TRANSPORT; OXYGEN-CONSUMPTION; GLUCOSE-METABOLISM; ORGANIC OSMOLYTES; PLASMA OSMOLALITY;
D O I：
10.1016/j.ygeno.2024.110833
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Myo-inositol is an important compatible osmolyte in vertebrates. This osmolyte is produced by the myo-inositol biosynthesis (MIB) pathway composed of myo-inositol phosphate synthase and inositol monophosphatase. These enzymes are among the highest upregulated proteins in tissues and cell cultures from teleost fish exposed to hyperosmotic conditions indicating high importance of this pathway for tolerating this type of stress. CRISPR/ Cas9 gene editing of tilapia cells produced knockout lines of MIB enzymes and control genes. Metabolic activity decreased significantly for MIB KO lines in hyperosmotic media. Trends of faster growth of the MIB knockout lines in isosmotic media and faster decline of MIB knockout lines in hyperosmotic media were also observed. These results indicate a decline in metabolic fitness but only moderate effects on cell survival when tilapia cells with disrupted MIB genes are exposed to hyperosmolality. Therefore MIB genes are required for full osmotolerance of tilapia cells.
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页数：12
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