Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production

F. Kent Hamra, Christopher T. Richie, Brandon K. Harvey

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Long Evans rat strains are applied as research models in a broad spectrum of biomedical fields (>15,800 citations, NCBI PubMed). Here, we report an approach to genetically modify the Long Evans rat germline in donor spermatogonial stem cells. Long Evans rat spermatogonial lines were derived from freshly isolated laminin-binding spermatogonia. Laminin-binding spermatogonia were cultured over multiple passages on fibroblast feeder layers in serum-free culture medium containing GDNF and FGF2. Long Evans rat spermatogonial lines were genetically modified by transposon transduction to express a germline, tdTomato reporter gene. Donor rat spermatogonial lines robustly regenerated spermatogenesis after transplantation into testes of busulfan-treated, allogenic, Long Evans rats. Donor-derived spermatogenesis largely restored testis size in the chemically sterilized, recipient Long Evans rats. Recipient Long Evans rats stably transmitted the tdTomato germline marker to subsequent generations. Overall, Long Evans rat spermatogonial lines provided effective donor germline vectors for genetically modifying Long Evans rats.

Original languageEnglish (US)
Pages (from-to)477-489
Number of pages13
JournalTransgenic Research
Volume26
Issue number4
DOIs
StatePublished - Aug 1 2017

Keywords

  • Allogeneic transplantation
  • Drug abuse
  • Drug addiction
  • Germline editing
  • Spermatogonial stem cells
  • Transgenic rats

ASJC Scopus subject areas

  • Biotechnology
  • Animal Science and Zoology
  • Agronomy and Crop Science
  • Genetics

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