CRISPR/Cas9-Mediated Transgenesis of the Masu Salmon (Oncorhynchus masou) elovl2 Gene Improves n-3 Fatty Acid Content in Channel Catfish (Ictalurus punctatus)

Omega-3 polyunsaturated fatty acids (n-3 PUFAs), particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), play a very important role in human health. Channel catfish ( Ictalurus punctatus ) is one of the leading freshwater aquaculture species in the USA, but has low...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2022-06, Vol.24 (3), p.513-523
Main Authors: Xing, De, Su, Baofeng, Li, Shangjia, Bangs, Max, Creamer, David, Coogan, Michael, Wang, Jinhai, Simora, Rhoda, Ma, Xiaoli, Hettiarachchi, Darshika, Alston, Veronica, Wang, Wenwen, Johnson, Andrew, Lu, Cuiyu, Hasin, Tasnuba, Qin, Zhenkui, Dunham, Rex
Format: Article
Language:English
Subjects:
Actin
Aquaculture
Biomedical and Life Sciences
Biosynthesis
Catfish
CRISPR
Deformation effects
Design optimization
Docosahexaenoic acid
Eicosapentaenoic acid
Engineering
Fatty acids
Fish
Freshwater
Freshwater & Marine Ecology
Freshwater aquaculture
Freshwater fishes
Gene editing
Gene expression
gRNA
Ictalurus punctatus
Inland water environment
Life Sciences
Microbiology
Morphometry
Muscles
Oncorhynchus masou
Original Article
Plasmids
Polyunsaturated fatty acids
Salmon
Tissue
Transgenic fish
Zoology
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Summary:Omega-3 polyunsaturated fatty acids (n-3 PUFAs), particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), play a very important role in human health. Channel catfish ( Ictalurus punctatus ) is one of the leading freshwater aquaculture species in the USA, but has low levels of EPA and DHA compared to some fish such as salmon. To improve EPA and DHA content, a modification of the n-3 PUFA biosynthetic pathway was achieved through the insertion of an elovl2 transgene isolated from masu salmon ( Oncorhynchus masou ) driven by a carp β-actin promoter using a two-hit by gRNA and two oligos with a targeting plasmid (2H2OP) CRISPR/Cas9 approach. Integration rate of the transgene was high (37.5%) and detected in twelve different tissues of P 1 transgenic fish with tissue-specific gene expression. Liver and muscle had relative high gene expression (13.4- and 9.2-fold change, respectively). Fatty acid analysis showed DHA content in the muscle from transgenic fish was 1.62-fold higher than in non-transgenic fish ( P  
ISSN:1436-2228
1436-2236
DOI:10.1007/s10126-022-10110-6