Testes of DAZL null neonatal sheep lack prospermatogonia but maintain normal somatic cell morphology and marker expression
| Autor: | Björn Oback, Zachariah McLean, Sarah Jane Appleby, Jingwei Wei, Russell G. Snell |
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| Rok vydání: | 2020 |
| Předmět: |
Male
0301 basic medicine Somatic cell Offspring Gene Expression Breeding Biology medicine.disease_cause Germline Animals Genetically Modified Andrology Mice 03 medical and health sciences DAZL 0302 clinical medicine Loss of Function Mutation Testis Genotype Genetics medicine Animals Cells Cultured Gene Editing Cloning Mutation Sheep 030219 obstetrics & reproductive medicine Base Sequence RNA-Binding Proteins Recombinational DNA Repair Cell Biology Fibroblasts Spermatogonia Phenotype 030104 developmental biology medicine.anatomical_structure Animals Newborn Biomarkers Germ cell Developmental Biology |
| Zdroj: | Molecular Reproduction and Development. 88:3-14 |
| ISSN: | 1098-2795 1040-452X |
| Popis: | Multiplying the germline would increase the number of offspring that can be produced from selected animals, accelerating genetic improvement for livestock breeding. This could be achieved by producing multiple chimaeric animals, each carrying a mix of donor and host germ cells in their gonads. However, such chimaeric germlines would produce offspring from both donor and host genotypes, limiting the rate of genetic improvement. To resolve this problem, we disrupted the RNA-binding protein DAZL and generated germ cell-deficient host animals. Using Cas9-mediated homology-directed repair (HDR), we introduced a DAZL loss-of-function mutation in male ovine fetal fibroblasts. Following manual single cell isolation, 4/48 (8.3%) of donor cell strains were homozygously HDR-edited. Sequence-validated strains were used as nuclear donors for somatic cell cloning to generate three lambs, which died at birth. All DAZL null male neonatal sheep lacked germ cells on histological sections and showed greatly reduced germ cell markers. Somatic cells within their testes were morphologically intact and expressed normal levels of lineage-specific markers, suggesting that the germ cell niche remained intact. This extends the DAZL mutant phenotype beyond mice into agriculturally relevant ruminants, providing a pathway for using absolute germline transmitters in rapid livestock improvement. |
| Databáze: | OpenAIRE |
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