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Application of genome editing 8:
Farm animals in aquatic systems
Anna Troedsson-Wargelius
Institute of Marine Research, Be...
Aquaculture environments for fish and shellfish
Ottinger et al 2016
Ottinger et al 2016
Global aquaculture production
Norwegian aquaculture of Atlantic salmon
• Norway is the world’s largest producer of Atlantic salmon
• > 1.2 million metri...
-Environmental impact
Disease
Genetic introgression of
farmed fish into wild populations
-Feed resources
Sustainability is...
Atlantic Salmon genome
Lien et al 2016
Ayllon et al 2015
Kjærner- Semb et al 2016
Establishment of CRISPR/Cas9 methodology in
Atlantic salmon
Edvardsen et al., Plos One (2014)
Injection of guide RNA + Cas...
Generation time in salmon 3-4 years
CRISPR/Cas9
Research tool and possible industrial applications
in salmon aquaculture
- Genetic containment of wild populat...
Environmental concerns
- Disease
- Genetic introgression of farmed escapees into wild populations
- Feed resources
Triploid sterile salmon
Genetic containment
- Produced by pressure treatment of eggs
- One extra set of chromosomes  ster...
Sterile salmon through germ cell ablation
Genetic containment
1) Broodstock/eggs:
Inhibit germ cell formation
2) Juveniles...
Sterile salmon through germ cell ablation
Genetic containment
χ
Germ cell-free (sterile) dnd-knockout
dnd + albino Knockout female Control female
Genetic containment Wargelius et al., Sc...
Germ cell-free salmon – what next?
Genetic containment Kleppe et al., Sci. Rep. (2017)
Ongoing: Long-term (2017-2020) grow...
Environmental concerns
- Disease
- Genetic introgression of farmed escapees into wild populations
- Feed resources
Omega-3 synthesis
Filet quality
EPA
DHA
Control
elovl2 + albino knockout
Datsmoor, Winge, in prep.
0
5
10
15
20
25
30
35
40
C18:3n3 C18:2n6 C20:2n6 C20:4n6 C20:5n3 C22:5n3 C22:6n3
%offattyacidsfromphospholipidsinthe
liver...
Environmental concerns
- Disease
- Genetic introgression of farmed escapees into wild populations
- Feed resources Number ...
Natura Disease resistance in wild strains
Kjærner-Semb et al 2016, BMC genomicsMove traits between strains
Gene editing by CRISPR/Cas9
NHEJ=
Non-Homologous End Joining
HDR=
Homology Directed Recombination
Region under selection for time of maturation
Move traits between strains Barson et al., Nature (2015)
Ayllon et al., Plos...
Ayllon et al., Plos Genetics (2015)Move traits between strains
SNPs explaining the trait in farmed fish
Straume, Edvardsen, Unpubl. res.
Move minor genetic elements between salmon
populations
Project:
AquaCrispr
Responsible Research Innovation (RRI)
- Interactions with public, presentations, media debates
- Ethics and integrity
-Loc...
Summary – Gene editing in aquaculture
- Targeted mutagenesis using Crispr-Cas9 is established
in a number of farmed fish s...
Reproduction and
Developmental biology group
at IMR
Anna Wargelius
Eva Andersson
Sven Leininger
Kai Ove Skaftnesmo
Geir La...
Thank your for the attention!
Farm animals in aquatic systems - Anna Troedsson-Wargelius
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Farm animals in aquatic systems - Anna Troedsson-Wargelius

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Farm animals in aquatic systems - Anna Troedsson-Wargelius

  1. 1. Application of genome editing 8: Farm animals in aquatic systems Anna Troedsson-Wargelius Institute of Marine Research, Bergen, Norway
  2. 2. Aquaculture environments for fish and shellfish Ottinger et al 2016
  3. 3. Ottinger et al 2016 Global aquaculture production
  4. 4. Norwegian aquaculture of Atlantic salmon • Norway is the world’s largest producer of Atlantic salmon • > 1.2 million metric tonnes in 2012- 2016 Norwegian farming of salmon, 1980-2015 What are the current challenges in salmon farming? Rainbow trout Atlantic salmon
  5. 5. -Environmental impact Disease Genetic introgression of farmed fish into wild populations -Feed resources Sustainability issues
  6. 6. Atlantic Salmon genome Lien et al 2016 Ayllon et al 2015 Kjærner- Semb et al 2016
  7. 7. Establishment of CRISPR/Cas9 methodology in Atlantic salmon Edvardsen et al., Plos One (2014) Injection of guide RNA + Cas9 in newly fertilized eggs Targeting a pigmentation gene: slc45a2 (albino) Successful editing = albinosSuccessful editing = F0 albinos Likewise the technology has been established in other aquaculture species.: Tilapia (Wang et al 2014) Cat fish (Zhong et al 2016) Carp (Khalil et al 2017)
  8. 8. Generation time in salmon 3-4 years
  9. 9. CRISPR/Cas9 Research tool and possible industrial applications in salmon aquaculture - Genetic containment of wild populations - Filet quality - Disease resistance - Move traits between strains
  10. 10. Environmental concerns - Disease - Genetic introgression of farmed escapees into wild populations - Feed resources
  11. 11. Triploid sterile salmon Genetic containment - Produced by pressure treatment of eggs - One extra set of chromosomes  sterile - Sensitive to warmer temperatures and low oxygen - More likely to develop skeletal deformities and cataract - Males og through puberty - Triplodization does not always result in 100% sterile fish batches - Prone to get bacterial disease
  12. 12. Sterile salmon through germ cell ablation Genetic containment 1) Broodstock/eggs: Inhibit germ cell formation 2) Juveniles: Induce apoptosis of germ cells
  13. 13. Sterile salmon through germ cell ablation Genetic containment χ
  14. 14. Germ cell-free (sterile) dnd-knockout dnd + albino Knockout female Control female Genetic containment Wargelius et al., Sci. Rep. (2016)
  15. 15. Germ cell-free salmon – what next? Genetic containment Kleppe et al., Sci. Rep. (2017) Ongoing: Long-term (2017-2020) growth experiment ~80 GCF and ~80 controls -Common garden -Induced early puberty -Measured growth and sex steroid levels -no puberty -normal growth
  16. 16. Environmental concerns - Disease - Genetic introgression of farmed escapees into wild populations - Feed resources
  17. 17. Omega-3 synthesis Filet quality EPA DHA Control elovl2 + albino knockout Datsmoor, Winge, in prep.
  18. 18. 0 5 10 15 20 25 30 35 40 C18:3n3 C18:2n6 C20:2n6 C20:4n6 C20:5n3 C22:5n3 C22:6n3 %offattyacidsfromphospholipidsinthe liver Wildtype Elovl2 knockout C20:5n3 C22:5n3 C22:6n3 p-values 0,0043 0,027 0,031 Filet quality Datsmoor, Winge, in prep. Effects of elovl2 knockout on DHA synthesis in the liver under standard diet
  19. 19. Environmental concerns - Disease - Genetic introgression of farmed escapees into wild populations - Feed resources Number of wild salmon that returns to rivers to breed
  20. 20. Natura Disease resistance in wild strains Kjærner-Semb et al 2016, BMC genomicsMove traits between strains
  21. 21. Gene editing by CRISPR/Cas9 NHEJ= Non-Homologous End Joining HDR= Homology Directed Recombination
  22. 22. Region under selection for time of maturation Move traits between strains Barson et al., Nature (2015) Ayllon et al., Plos Genetics (2015)
  23. 23. Ayllon et al., Plos Genetics (2015)Move traits between strains SNPs explaining the trait in farmed fish
  24. 24. Straume, Edvardsen, Unpubl. res. Move minor genetic elements between salmon populations Project: AquaCrispr
  25. 25. Responsible Research Innovation (RRI) - Interactions with public, presentations, media debates - Ethics and integrity -Local highschool integration (Nordahl Grieg og Amalie Skram)
  26. 26. Summary – Gene editing in aquaculture - Targeted mutagenesis using Crispr-Cas9 is established in a number of farmed fish species - F0 mutant studies are enabled - Homologous recombination of single causative SNPs is being explored (Disease resistance, control of reproduction, nutritional content, fish robustness) - Mutants representing sustainable traits are being studied (Germ cell-free fish, altered omega-3 metabolism, time of maturation etc.)
  27. 27. Reproduction and Developmental biology group at IMR Anna Wargelius Eva Andersson Sven Leininger Kai Ove Skaftnesmo Geir Lasse Taranger Erik Kjærner Semb Tomasz Furmanek Per Gunnar Fjelldal Thomas Fraser Tom Hansen Birgitta Norberg Hilal Güralp Anne Hege Straume Rolf Edvardsen Population genetics group at IMR Kevin Glover Fernando Ayllon Vidar Wennevik Calle Rubin Dorothy Dankel Alex Datsomoor Per Winge Nina Santi Maren Mommens Helge Tveiten Øivind Andersen Rüdiger W. Schulz Hildre Sindre Acknowledgements Funding: -Norwegian Research Council BIOTEK2021 project SALMOSTERILE HAVBRUK_BIOTEK2021 project SALMAT HAVBRUK project STERWELL and MATGEN -EU AQUACRISPR
  28. 28. Thank your for the attention!

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