Novel method for mass producing genetically sterile fish from surrogate broodstock via spermatogonial transplantation†

Nagasawa, Kazue; Ishida, Masato; Octavera, Anna; Kusano, Kazunari; Kezuka, Fumi; Kitano, Takeshi; Yoshiura, Yasutoshi; Yoshizaki, Goro

doi:10.1093/biolre/ioy204

Cited by 34 publications

(16 citation statements)

References 41 publications

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“…However, in contrast to other non-transgenic methods like embryo immersion in dnd Vivo-conjugated morpholino 44 or triploidization 10 , our approach if functional in all steps display a significant advantage since it will always produce 100% sterility in combination with the transmissibility of sterility to the next generation. One of the significant advantages of this method is that it can be applied to both sexes without additional steps such as germ cell transplantation or sex reversal 45 . The long reproductive cycle makes the implementation and standardization of the procedure time-dependent; however, after creating the broodstock, microinjections of dnd mRNA in dnd crispant offspring will enable further restocking.…”

Section: Discussionmentioning

confidence: 99%

Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon

Güralp

Skaftnesmo

Kjærner-Semb

et al. 2020

Sci Rep

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Genetic introgression of escaped farmed Atlantic salmon (Salmo salar) into wild populations is a major environmental concern for the salmon aquaculture industry. Using sterile fish in commercial aquaculture operations is, therefore, a sustainable strategy for bio-containment. So far, the only commercially used methodology for producing sterile fish is triploidization. However, triploid fish are less robust. A novel approach in which to achieve sterility is to produce germ cell-free salmon, which can be accomplished by knocking out the dead-end (dnd) gene using CRISPR-Cas9. The lack of germ cells in the resulting dnd crispants, thus, prevents reproduction and inhibits subsequent large-scale production of sterile fish. Here, we report a rescue approach for producing germ cells in Atlantic salmon dnd crispants. To achieve this, we co-injected the wild-type (wt) variant of salmon dnd mRNA together with CRISPR-Cas9 constructs targeting dnd into 1-cell stage embryos. We found that rescued one-year-old fish contained germ cells, type A spermatogonia in males and previtellogenic primary oocytes in females. The method presented here opens a possibility for large-scale production of germ-cell free Atlantic salmon offspring through the genetically sterile broodstock which can pass the sterility trait on the next generation.

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Section: Discussionmentioning

confidence: 99%

Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon

Güralp

Skaftnesmo

Kjærner-Semb

et al. 2020

Sci Rep

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“…This, however, is not a practical method because gametes cannot be obtained from sterile individuals produced by these methods and microinjection into the fertilized egg is required every time to produce sterile fish. Nagasawa et al (2018) developed a method to mass produce sterile individuals using surrogate broodstock. Oogenesis of fish generally progresses when ovaries receive stimulation from follicle-stimulating hormone (FSH) secreted from the pituitary gland (Clelland and Peng 2009).…”

Section: Producing Sterile Fishmentioning

confidence: 99%

“…When we fertilized these eggs with sperm of a pseudo-male of a FSH-receptor-mutant medaka (masculinized XX genotype medaka), all of the next generation was XX (and all-sterile females). Thus, it was possible to mass produce sterile individuals with disrupted FSH receptor genes (Nagasawa et al 2018). In principle, this method can be applied to a wide variety of aquaculture fish species.…”

Section: Producing Sterile Fishmentioning

confidence: 99%

Application of surrogate broodstock technology in aquaculture

Yoshizaki

Yazawa

2019

Fish Sci

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Surrogate broodstock technology facilitates the production of donor-derived gametes in surrogates, and comprises transplanting germ cells of a donor into recipients of a different strain or different species. The following applications of this technology are expected in the field of aquaculture: (1) the efficient and reliable production of offspring carrying superior genetic traits by transplanting donor germ cells from a single selected fish with superior traits into many recipient fish; (2) the reduction of the time required to breed fish by using a recipient species with a short generation time to produce gametes of a species with a long generation time; (3) the long-term storage of valuable species or strains as genetic resources by cryopreserving germ cells for transplantation; (4) the mass production of genetically sterile fish by transplanting germ cells of a donor fish that is sterile due to a mutation in the somatic cells into normal recipients without this mutation. It is expected that a combination of these techniques will greatly accelerate the breeding of aquaculture species. It is important to adapt surrogate broodstock technology to a wider range of fishery species and further improve the efficiency of donor-derived gamete production when using surrogate broodstock.

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“…On the other hand it is necessary to mention, that genomic resources are necessary to target dnd gene essential for PGCs migration by knock down or knock out techniques [42]. Hybridisation of common carp or gold sh might be another potential tool as hybrids were used successfully as surrogates in different species [46,47]. Performed studies reported sterility in cross of common carp a crucian carp [48] but no data are available regarding sex ratio of sterile hybrids.…”

Section: Discussionmentioning

confidence: 99%

Production of common carp donor-derived offspring from goldfish surrogate broodstock

Franěk

Kašpar

Gela

et al. 2020

Preprint

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Background: Common carp is the fourth most-produced species in worldwide aquaculture. Significant efforts are invested in breeding and preservation of genetic integrity of this important species. However, maintaining carp gene bank in situ can be considered as demanding due to its big body size. Recent progress in reproductive biotechnologies in fish allows improving some unfavourable characteristics of a target species using surrogate reproduction. Germ stem cells (gamete precursors) from one species are transplanted into different surrogate species with small body size. After maturation, surrogates are producing donor-derived progeny. Efficient protocols for cryopreservation of carp male and female germ stem cells have been developed lately. Thus, the next logical goal was to assess the potential of goldfish surrogate to produce donor-derived gametes of common carp after intraperitoneal transplantation of testicular cells. Results: High transplantation success was achieved when 44% of the surviving goldfish produced pure donor-derived gametes of common carp. More importantly, both viable eggs and sperm giving rise to pure common carp progeny were produced, witnessing sustainability of the presented method. Donor-derived identity of the offspring was confirmed by genotyping and typical phenotype corresponding to the donor species. Reproductive performance of chimeras was similar to goldfish controls. Assessment of gamete characteristics showed that the size of donor-derived eggs is between control carp and goldfish eggs. Interestingly, flagellum length in donor-derived spermatozoa was comparable to common carp flagellum and significantly shorter than goldfish flagellum. Conclusions: In this study, we succeeded in the production of pure common carp progeny from surrogate goldfish recipients transplanted intraperitoneally by testicular germ cells. Here we reported production of viable eggs between most distant species up to date. Good reproductive performance of goldfish germline chimeras gives a promising prospect for further analysis about the long-term reproductive performance of surrogates, recovery of cryopreserved germ cells or production of monosex stocks. Presented technology is ready to ease needs for carp breeds preservation and their recovery using many times smaller goldfish surrogates.

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Novel method for mass producing genetically sterile fish from surrogate broodstock via spermatogonial transplantation†

Cited by 34 publications

References 41 publications

Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon

Rescue of germ cells in dnd crispant embryos opens the possibility to produce inherited sterility in Atlantic salmon

Application of surrogate broodstock technology in aquaculture

Production of common carp donor-derived offspring from goldfish surrogate broodstock

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