B6J.Cg-Oip5os1^tm1.2Dpbl/J

Stock number: 033986
Product name: Cyrano^-; Cyrano KO
Research areas: Cell Biology Research

Description

Cyrano KO mice carry a knock-out of the mouse Oip5os1 (Opa interacting protein 5, opposite strand 1; also called Cyrano) locus. Homozygotes demonstrate increased levels of miR-7 (microRNA 7). This strain is useful in studies of gene regulation by non-coding RNAs (ncRNAs).

Detailed description

Oip5os1 (Opa interacting protein 5, opposite strand 1; also called Cyrano) expresses a long intergenic non-coding RNA (ncRNA) that directs the effective degradation of miR-7 (microRNA 7). Loss of miR-7 prevents the repression of mRNAs targeted by miR-7, and enables the accumulation of Cdr1os (cerebellar degeneration related antigen 1, opposite strand; also called Cdr1as), a circular RNA known to regulate neuronal activity.

These Cyrano KO mice carry a targeted knock-out of the mouse Oip5os1 locus generated by crossing Cyrano CKO floxed mice (Stock No. 033987) with animals expressing CMV-cre (Stock No. 006054). RNA sequencing confirms that the deletion of the first half of exon 3, which includes the most conserved region of Cyrano associated with extensive miR-7 pairing, results in the complete loss of the targeted region as well as more than 90% of flanking Cyrano RNA. Homozygotes are born at expected Mendelian frequency and show no overt abnormalities. Cyrano loss does not seem to affect performance in standard neurobehavioral tests.

Homozygotes demonstrate increased levels of miR-7 microRNA in many tissues. miR-7a (Mir7-1) and miR-7b (Mir7b) levels are increased by 40- and 47-fold in cerebellum and 6- and 7-fold in hippocampus.

Development

A loxP site was introduced to intron 2 and an FRT-neomycin-FRT-loxP cassette was placed in exon 3 of the mouse Oip50s1 locus via homologous recombination in 129S6/SvEvTac x C57BL/6J-derived embryonic stem (ES) cells. Resulting chimeric mice were bred with FLPe transgenic mice (Stock No. 003946) to excise the FRT-flanked neomycin cassette, producing the Cyrano^fl allele in which the 5' portion of exon 3 is flanked by loxP sites.

Cyrano^fl mice (Stock No. 033987) were bred to CMV-cre mice (Stock No. 006054) to generate this constitutive knock-out of the mouse Oip5os1 gene (Cyrano^-). This line was backcrossed to C57BL/6J for at least 8 generations by the donating laboratory.

Allele

Symbol: Oip5os1^tm1.2Dpbl
Name: targeted mutation 1.2, David P Bartel
MGI accession ID: MGI:6295970
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Oip5os1
Marker name: Opa interacting protein 5, opposite strand 1
Marker synonyms: 1190006L01Rik; 1700020I14Rik; 4933437I03Rik; Cyrano; linc-oip5; Oip5as1
Chromosome: 2
Strain of origin: 129S6/SvEvTac x C57BL/6J
Molecular note: A loxP site was inserted upstream of exon 3. An FRT-flanked neomycin resistance cassette with a 3' loxP site was inserted in exon 3 downstream of the site for induction of Mir 7 degradation. Flp-mediated recombination removed the selection cassette. Cre-mediated recombination removed the coding sequence of exon 3 up to and including the Mir site.