STOCK Rab11a^tm1.1Ngao/J

Stock number: 024110
Product name: Rab11a^fl
Research areas: Cancer Research, Cell Biology Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

The Rablla^fl allele has loxP sites flanking exons 3-4 of the Rab11a gene. These mice allow generation of tissue-specific Rablla knockout, and may be useful for studying the function of this recycling endosome resident small GTPase in intracellular trafficking, Toll-like receptors (TLRs), inflammatory responses and intestinal host-microbial homeostasis.

Detailed description

The Rablla^fl allele has loxP sites flanking exons 3-4 of the Rab11a gene. The Rab11a gene encodes a RAS oncogene family protein that functions as a small GTPase in intracellular membrane trafficking, and is also a component of the recycling endosome. Mice homozygous for the Rablla^fl allele are viable and fertile with no reported abnormalities. When bred to mice that express Cre recombinase, the resulting offspring may be useful in generating tissue-specific Rablla knockout.

For example, when bred to germline Cre-expressing mice, the resulting Rab11a global knockout homozygotes die in utero around the implantation stage. When Rablla^fl mice are bred to Villin-Cre transgenic mice (Stock Nos. 004586 / 018963), the resulting Rablla^ΔIEC homozygotes lack Rablla expression in intestinal intraepithelial cells, and exhibit epithelial cell-intrinsic cytokine production, inflammatory bowel phenotype and early mortality.

Development

The targeting construct was designed by Dr. Nan Gao (Rutgers University) to insert a loxP site upstream of exon 3, and a frt-flanked PGK-neo cassette followed by a second loxP site just downstream of exon 4 of the Rab11a gene on chromosome 9.

The donating investigator confirms that the construct was electroporated into 129S4/SvJae-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were were identified and injected into recipient blastocysts. Chimeric males were bred to C57BL/6J females for germline transmission. The mice were then bred to ACT-FLPe animals (Stock No. 003800; B6;SJL-Tg(ACTFLPe)9205Dym/J) to delete frt-flanked PGK-neo; resulting in mice with the Rablla^fl allele.

The Rablla^fl colony was maintained on this mixed genetic background for several generations (and the Flp-expressing transgene was removed) prior to sending males (with agouti coat color) to The Jackson Laboratory Repository in 2015.

Upon arrival, males were used to cryopreserve sperm. To establish our living mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

Allele

Symbol: Rab11a^tm1.1Ngao
Name: targeted mutation 1.1, Nan Gao
MGI accession ID: MGI:5608786
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Rab11a
Marker name: RAB11A, member RAS oncogene family
Chromosome: 9
Strain of origin: 129S4/SvJae
Molecular note: A loxP site was inserted at 242 bp downstream of the second exon and a and a FRT-flanked PGK-neo cassette followed by a second loxP site was introduced at 210 bp downstream of the fourth exon. Flp-mediated recombination removed the FRT-flanked neo cassette.