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Testes of DAZL null sheep lack spermatogonia and maintain normal somatic cells

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posted on 2023-05-03, 21:25 authored by Zach McLean, Sarah ApplebySarah Appleby, Jingwei WeiJingwei Wei, Russell Snell, Björn ObackBjörn Oback
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 and produce chimaeras with absolute donor germline transmission, we have 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. Somatic cells within their testes were morphologically intact and expressed normal levels of somatic cell-specific marker genes, indicating 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 transmitters in rapid livestock improvement.

History

Rights statement

The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.

Publication date

2019-11-29

Project number

  • Non revenue

Language

  • English

Does this contain Māori information or data?

  • No

Publisher

bioRxiv

Citation

McLean, Z., Appleby, S. J., Wei, J., Snell, R. G., & Oback, B. (2019). Testes of DAZL null sheep lack spermatogonia and maintain normal somatic cells, bioRxiv. doi:10.1101/848036

Funder

Ministry of Business Innovation & Employment

Contract number

A24261

Job code

15795

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