B6.Cg-Tg(RP24-131B16/EGFP)13Ghan/J

Stock number: 022086
Research areas: Research Tools

Description

These transgenic mice express tagged antisense piRNAs from a Chromosome 17 cluster during male meiosis.

Detailed description

Piwi-interacting RNAs (piRNAs), a discrete class of small RNAs, are expressed in mammalian germline cells and play a critical role in germline development. Arising from discrete genomic loci (clusters), they are selected from long precursor transcripts through unknown mechanisms to guard germ cell genomes against the activity of mobile genetic elements.

This BAC-derived transgene encodes approximately 100 kb of a mouse Chromosome 17 piRNA cluster (mm9 assembly Chr. 17:27427600-27488899) and incorporates a GFP marker inserted antisense to the direction of transcription. Expression of the transgene recapitulates that of the endogenous locus, with transcription starting in male germ cells post-puberty. GFP-tagged antisense piRNAs are produced during male meiosis. No sense GFP is expressed, thus no fluorescence can be detected.

This transgenic strain demonstrates that piRNA clusters can be separated from their native genomic locations and contain sufficient information to funnel their RNA products into the piRNA biogenesis pathway.

No phenotypic differences between founder lines 13 and 37 (see Stock No. 019494) have been noted.

Development

BAC clone RP24-131B16 containing a mouse Chromosome 17 piRNA cluster was modified to express FRT-PGK-gb2-neo-FRT and EGFP (inserted antisense) from the 3' end of the cluster downstream of a central bi-directional promoter. The transgenic vector was introduced to B6SJLF1 hybrid oocytes. The donating investigator reports that this strain was backcrossed to C57BL/6 for approximately 10 generations (see SNP note below). This is founder line 13.

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 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. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, all 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 C57BL/6N genetic background.

Allele

Symbol: Tg(RP24-131B16/EGFP)13Ghan
Name: transgene insertion 13, Gregory J Hannon
MGI accession ID: MGI:5471221
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(RP24-131B16/EGFP)13Ghan
Marker name: transgene insertion 13, Gregory J Hannon
Chromosome: UN
Strain of origin: (C57BL/6 x SJL)F1
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: Expression of the transgene recapitulates that of the endogenous locus, with transcription starting in male germ cells post-puberty.
Molecular note: BAC clone RP24-131B16 containing a mouse Chromosome 17 piRNA cluster was modified to express FRT-PGK-gb2-neo-FRT and EGFP (inserted antisense) from the 3' end of the cluster downstream of a central bi-directional promoter. A line was establsiehd from founder 13.