STOCK Tg(Nes-EGFP)33Enik/J

Stock number: 033927
Product name: nestin-GFP

Strain synonyms: NGFP
Research areas: Neurobiology Research, Research Tools

Description

Nestin-GFP mice express an enhanced green fluorescent protein (EGFP) directed to the neuroepithelium of the embryonic brain using the regulatory elements of the Nes (nestin) gene. This mutant mouse strain may be useful in visualizing neural stem and progenitor cells in embryonic and adult brains.

Detailed description

The nestin-GFP transgene uses the mouse Nes (nestin) regulatory elements to direct expression of the enhanced green fluorescent protein (EGFP) to the neuroepithelium of the embryonic brain. Nes encodes an intermediate filament protein expressed in the neural stem cells of developing and adult brains. EGFP expression mimics the endogenous promoter/enhancer expression pattern in neural tissue. EGFP expression is observed as early as embryonic day 7. Signal intensity is greatest between E14 and E15, by E20 EGFP signal is seen in the narrow area of subbventricular zone. In the postnatal brain, EGFP expression is observed in the cerebellum, and gradually becomes limited to the Bergmann glial cells. Expression is absent in differentiated neurons. Mice homozygous for the transgene are viable and fertile. This mutant mouse strain may be useful in visualizing neural stem and progenitor cells in embryonic and adult brains.

Development

The transgenic construct contains a 5.8 kb fragment of the Nes (nestin) promoter, an enhanced green fluorescent protein (EGFP) followed by an SV40 polyA and a 1.8 kb fragment containing the Nes enhancer region in the second intron. The inclusion of the enhancer from the second intron directs expression to neural tissues. The linearized construct was microinjected into fertilized C57BL/6J x BALB/cByJ hybrid oocytes. Founder line 33 was subsequently established. Founder animals were bred to C57BL/6J female mice and then backcrossed for at least five generations by the donating lab (see SNP note below). Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.



In 2019 a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers throughout the genome, on Chromosome 17 was segregating with BALB/cByJ, and another marker on Chromosome 17 was segregating with an unknown source. Also, 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Tg(Nes-EGFP)33Enik
Name: transgene insertion 33, Grigori Enikolopov
MGI accession ID: MGI:5523870
Generation method: Transgenic
Attributes: Reporter
Synonyms: nestin-GFP
Marker symbol: Tg(Nes-EGFP)33Enik
Marker name: transgene insertion 33, Grigori Enikolopov
Chromosome: UN
Strain of origin: (C57BL/6 x BALB/cBy)F1
Site of expression: neuroepithelium of the embryonic brain
Molecular note: The transgene contains an EGFP and polyadenylation sequence inserted between the neural stem cell-specific promoter and the enhancer containing second intron. Three lines were generated, including line 33.