PUBLICATION
Application of dead end-knockout zebrafish as recipients of germ cell transplantation
- Authors
- Li, Q., Fujii, W., Naito, K., Yoshizaki, G.
- ID
- ZDB-PUB-170722-1
- Date
- 2017
- Source
- Molecular reproduction and development 84(10): 1100-1111 (Journal)
- Registered Authors
- Keywords
- gene knockout, spermatogonia, surrogate broodstock, zinc finger nuclease
- MeSH Terms
-
- Adult Germline Stem Cells/cytology
- Adult Germline Stem Cells/transplantation
- Animals
- Animals, Genetically Modified
- Cell Transplantation/methods*
- Embryo, Nonmammalian
- Gene Knockout Techniques/methods
- Germ Cells/transplantation*
- Male
- Models, Animal*
- RNA-Binding Proteins/genetics*
- Transplant Recipients*
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/physiology*
- Zebrafish Proteins/genetics*
- PubMed
- 28731265 Full text @ Mol. Reprod. Dev.
Citation
Li, Q., Fujii, W., Naito, K., Yoshizaki, G. (2017) Application of dead end-knockout zebrafish as recipients of germ cell transplantation. Molecular reproduction and development. 84(10):1100-1111.
Abstract
Germ cell transplantation is a promising technology for the propagation of endangered or valuable fishes. In this technique, sterile male and female recipient fish are injected with donor germ cells so they can produce viable gametes derived from the donor cells. The dead end (dnd) gene is involved in the migration of primordial germ cells; therefore, dnd-knockout zebrafish are expected to be germ-cell-free, making them suitable recipients for germ cell transplantation. dnd mutants were produced by microinjecting 2 nl of 10 ng/μl cRNAs encoding zinc finger nucleases against dnd into the blastodisc of zebrafish embryos before the cell cleavage stage. One of the resulting founder males was mated with a wild-type female, and produced heterozygous mutants in the F1 generation. Mating of these F1 mutants produced an F2 generation with approximately 25% of the clutch being homozygous mutant (dnd-knockout), male, and lacking germ cells (as confirmed by expression analyses of vasa). The resulting dnd-knockout zebrafish males were tested for suitability as germ cell transplantation recipients by intraperitoneal transplantation of testicular cells prepared from vasa-gfp zebrafish. GFP-positive germ cells incorporated into the germ cell-free gonads of the dnd-knockout recipients matured into functional sperm. Progeny tests revealed that the sperm from these dnd-knockout recipients were derived entirely from donor cells. Thus, we demonstrated that homozygous dnd mutants became germ cell-free males that are able to nurse donor-derived germ cells. This article is protected by copyright. All rights reserved.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping