A transgenic DND1GFP fusion allele reports in vivo expression and RNA-binding targets in undifferentiated mouse germ cells†.

Journal Article (Journal Article)

In vertebrates, the RNA-binding protein (RBP) dead end 1 (DND1) is essential for primordial germ cell (PGC) survival and maintenance of cell identity. In multiple species, Dnd1 loss or mutation leads to severe PGC loss soon after specification or, in some species, germ cell transformation to somatic lineages. Our investigations into the role of DND1 in PGC specification and differentiation have been limited by the absence of an available antibody. To address this problem, we used CRISPR/Cas9 gene editing to establish a transgenic mouse line carrying a DND1GFP fusion allele. We present imaging analysis of DND1GFP expression showing that DND1GFP expression is heterogeneous among male germ cells (MGCs) and female germ cells (FGCs). DND1GFP was detected in MGCs throughout fetal life but lost from FGCs at meiotic entry. In postnatal and adult testes, DND1GFP expression correlated with classic markers for the premeiotic spermatogonial population. Utilizing the GFP tag for RNA immunoprecipitation (RIP) analysis in MGCs validated this transgenic as a tool for identifying in vivo transcript targets of DND1. The DND1GFP mouse line is a novel tool for isolation and analysis of embryonic and fetal germ cells, and the spermatogonial population of the postnatal and adult testis.

Full Text

Duke Authors

Cited Authors

  • Ruthig, VA; Yokonishi, T; Friedersdorf, MB; Batchvarova, S; Hardy, J; Garness, JA; Keene, JD; Capel, B

Published Date

  • April 1, 2021

Published In

Volume / Issue

  • 104 / 4

Start / End Page

  • 861 - 874

PubMed ID

  • 33394034

Pubmed Central ID

  • PMC8324984

Electronic International Standard Serial Number (EISSN)

  • 1529-7268

Digital Object Identifier (DOI)

  • 10.1093/biolre/ioaa233

Language

  • eng

Conference Location

  • United States