Indel locations are determined by template polarity in highly efficient in vivo CRISPR/Cas9-mediated HDR in Atlantic salmon

AbstractThe in vivo functions of Atlantic salmon fatty acyl desaturases (fads2), Δ6fads2-a, Δ6fads2-b, Δ6fads2-c and Δ5fads2 in long chain polyunsaturated fatty acid (LC-PUFA) synthesis in salmon and fish in general remains to be elucidated. Here, we investigate in vivo functions and in vivo functional redundancy of salmon fads2 using two CRISPR-mediated partial knockout salmon, Δ6abc/5Mt with mutations in Δ6fads2-a, Δ6fads2-b, Δ6fads2-c and Δ5fads2, and Δ6bcMt with mutations in Δ6fads2-b and Δ6fads2-c. F0 fish displaying high degree of gene editing (50–100%) were fed low LC-PUFA and high LC-PUFA diets, the former containing reduced levels of eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids but higher content of linoleic (18:2n-6) and alpha-linolenic (18:3n-3) acids, and the latter containing high levels of 20:5n-3 and 22:6n-3 but reduced compositions of 18:2n-6 and 18:3n-3. The Δ6abc/5Mt showed reduced 22:6n-3 levels and accumulated Δ6-desaturation substrates (18:2n-6, 18:3n-3) and Δ5-desaturation substrate (20:4n-3), demonstrating impaired 22:6n-3 synthesis compared to wildtypes (WT). Δ6bcMt showed no effect on Δ6-desaturation compared to WT, suggesting Δ6 Fads2-a as having the predominant Δ6-desaturation activity in salmon, at least in the tissues analyzed. Both Δ6abc/5Mt and Δ6bcMt demonstrated significant accumulation of Δ8-desaturation substrates (20:2n-6, 20:3n-3) when fed low LC-PUFA diet. Additionally, Δ6abc/5Mt demonstrated significant upregulation of the lipogenic transcription regulator, sterol regulatory element binding protein-1 (srebp-1) in liver and pyloric caeca under reduced dietary LC-PUFA. Our data suggest a combined effect of endogenous LC-PUFA synthesis and dietary LC-PUFA levels on srebp-1 expression which ultimately affects LC-PUFA synthesis in salmon. Our data also suggest Δ8-desaturation activities for salmon Δ6 Fads2 enzymes.

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Oral Administration of Lactococcus lactis Producing Interferon Type II, Enhances the Immune Response Against Bacterial Pathogens in Rainbow Trout

Frontiers in Immunology ◽

10.3389/fimmu.2021.696803 ◽

2021 ◽

Vol 12 ◽

Author(s):

Alvaro Santibañez ◽

Diego Paine ◽

Mick Parra ◽

Carlos Muñoz ◽

Natalia Valdes ◽

...

Keyword(s):

Immune Response ◽

Lactic Acid ◽

Rainbow Trout ◽

Lactic Acid Bacteria ◽

Atlantic Salmon ◽

Oral Administration ◽

Bacterial Infection ◽

Type Ii ◽

Recombinant Interferon

Lactic acid bacteria are a powerful vehicle for releasing of cytokines and immunostimulant peptides at the gastrointestinal level after oral administration. However, its therapeutic application against pathogens that affect rainbow trout and Atlantic salmon has been little explored. Type II interferon in Atlantic salmon activates the antiviral response, protecting against viral infection, but its role against bacterial infection has not been tested in vivo. In this work, through the design of a recombinant lactic acid bacterium capable of producing Interferon gamma from Atlantic salmon, we explore its role against bacterial infection and the ability to stimulate systemic immune response after oral administration of the recombinant probiotic. Recombinant interferon was active in vitro, mainly stimulating IL-6 expression in SHK-1 cells. In vivo, oral administration of the recombinant probiotic produced an increase in IL-6, IFNγ and IL-12 in the spleen and kidney, in addition to stimulating the activity of lysozyme in serum. The challenge trials indicated that the administration of the IFNγ-producing probiotic doubled the survival in fish infected with F. psychrophilum. In conclusion, our results showed that the oral administration of lactic acid bacteria producing IFNγ managed to stimulate the immune response at a systemic level, conferring protection against pathogens, showing a biotechnological potential for its application in aquaculture.

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A novel in vivo model for rapid evaluation of Aliivibrio salmonicida infectivity in Atlantic salmon

Aquaculture ◽

10.1016/j.aquaculture.2013.10.025 ◽

2014 ◽

Vol 420-421 ◽

pp. 112-118 ◽

Cited By ~ 9

Author(s):

Alexander Kashulin ◽

Henning Sørum

Keyword(s):

Atlantic Salmon ◽

In Vivo Model ◽

Rapid Evaluation ◽

Aliivibrio Salmonicida

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Tumor Regression: Injectable Smart Phase-Transformation Implants for Highly Efficient In Vivo Magnetic-Hyperthermia Regression of Tumors (Adv. Mater. 44/2014)

Advanced Materials ◽

10.1002/adma.201470305 ◽

2014 ◽

Vol 26 (44) ◽

pp. 7532-7532

Author(s):

Yu Chen ◽

Ling Jiang ◽

Ronghui Wang ◽

Ming Lu ◽

Qunxia Zhang ◽

...

Keyword(s):

Phase Transformation ◽

Tumor Regression ◽

Magnetic Hyperthermia ◽

Highly Efficient

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Novel cyclometalated iridium(iii) phosphine-imine (P^N) complexes: highly efficient anticancer and anti-lung metastasis agents in vivo

Inorganic Chemistry Frontiers ◽

10.1039/c9qi01492f ◽

2020 ◽

Vol 7 (5) ◽

pp. 1273-1283 ◽

Cited By ~ 2

Author(s):

Zhishan Xu ◽

Yuliang Yang ◽

Xianglei Jia ◽

Lihua Guo ◽

Xingxing Ge ◽

...

Keyword(s):

Lung Metastasis ◽

Tumour Growth ◽

Combined Treatment ◽

Highly Efficient

Iridium(iii)-based complexes with phosphine-imine (P^N) ligands are synthesized and authenticated. The combined treatment with Ir(iii) and BIX01294 potently inhibited tumour growth and lung metastasis in vitro and in vivo.

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IN VIVO METABOLISM OF STEROID HORMONES BY SOCKEYE SALMON: (A) IMPAIRED HORMONE CLEARANCE IN MATURE AND SPAWNED PACIFIC SALMON (O. NERKA) (B) PRECURSORS OF 11-KETOTESTOSTERONE

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o63-100 ◽

1963 ◽

Vol 41 (4) ◽

pp. 875-887 ◽

Cited By ~ 47

Author(s):

D. R. Idler ◽

B. Truscott ◽

H. C. Freeman ◽

V. Chang ◽

P. J. Schmidt ◽

...

Keyword(s):

Atlantic Salmon ◽

Sockeye Salmon ◽

Pacific Salmon ◽

In Vivo Metabolism ◽

Hormone Metabolism ◽

The Pacific ◽

Imminent Death ◽

High Concentrations ◽

Sexually Mature

Intra-arterially injected cortisone-4-C14 and cortisol-4-C14 were cleared from the plasma of sexually mature and spawned sockeye salmon (O. nerka) at a much slower rate than from the plasma of immature sockeye and spawned Atlantic salmon (S. salar). The results explain the elevated hormone levels found in the blood of mature and spawned sockeye salmon. The normal clearance rate found with Atlantic salmon, which frequently survive spawning, would indicate that the impaired hormone metabolism was associated with the imminent death of the Pacific salmon rather than with the act of spawning.Testosterone and 17α-hydroxyprogesterone were found to be precursors of 11-ketotestosterone, a sex hormone found in high concentrations in the blood of mature sockeye salmon. Testosterone was also formed in vivo from 17α-hydroxyprogesterone. The results suggest more than one pathway for the synthesis of 11-ketotestosterone in salmon. Cortisol was converted to cortisone but no conversion of the former to 11-ketotestosterone could be demonstrated.

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