STOCK Tg(Nanog-HIST1H2BB/EGFP)38Hadj/J

Stock number: 027563
Product name: Nanog:H2B-GFP
Research areas: Research Tools

Description

Nanog:H2B-GFP transgenic mice may be useful for single-cell resolution quantitative visualization of the pluripotent epiblast lineage in vivo.

Detailed description

Nanog:H2B-GFP transgenic mice express an H2B-EGFP fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] followed C-terminally by Enhanced Green Fluorescent Protein gene) under the control of the murine Nanog homeobox (Nanog) promoter. Mice homozygous for the transgene are viable and fertile. NANOG is a transcription factor normally expressed in embryonic stem cells (ESCs) and functions as a key factor in maintaining pluripotency. Nanog:H2B-GFP transgenic mice do not express Nanog from the transgene, making accurate quantitative readout of endogenous Nanog expression possible. Specifically, GFP expression is evident in pluripotent epiblast cells in embryos at the 8–16 cell stage, with higher levels of expression eventually restricted to the pluripotent epiblast seen in early and mid-blastocysts. Irreversible commitment to pluripotent epiblast lineage can be visualized in these reporter mice.

Development

The Nanog:H2B-GFP transgene was designed with the murine Nanog homeobox (Nanog) promoter driving expression of an H2B-EGFP fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] with a C-terminal Enhanced Green Fluorescent Protein gene), followed by an internal ribosomal entry site (IRES), a puromycin resistance cassette, and simian virus 40 (SV40) polyadenylation site. The transgene was microinjected into CBA/J x C57BL/6J fertilized oocytes, and mice from founder line number 38 were bred to outbred ICR mice to establish a colony. Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.

Allele

Symbol: Tg(Nanog-HIST1H2BB/EGFP)38Hadj
Name: transgene insertion 38, Anna-Katerina Hadjantonakis
MGI accession ID: MGI:5634139
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Nanog-HIST1H2BB/EGFP)38Hadj
Marker name: transgene insertion 38, Anna-Katerina Hadjantonakis
Chromosome: UN
Strain of origin: CBA/J x C57BL/6J
Expressed genes: H2BC3,H2B clustered histone 3,human; GFP,Green Fluorescent Protein,
Site of expression: H2B-EGFP fusion protein expression occurs in pluripotent epiblast cells in embryos.
Molecular note: The transgene is designed with the murine Nanog promoter driving expression of an H2B-EGFP fusion protein (coding sequence for the human HIST1H2BB gene [histone 1 H2bb] with a C-terminal Enhanced Green Fluorescent Protein gene), followed by an internal ribosomal entry site (IRES), a puromycin resistance cassette, and simian virus 40 (SV40) polyadenylation site.