The Rabllafl 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.
Nan Gao, Rutgers University
The Rabllafl 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 Rabllafl 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 Rabllafl mice are bred to Villin-Cre transgenic mice (Stock Nos. 004586 / 018963), the resulting RabllaΔIEC homozygotes lack Rablla expression in intestinal intraepithelial cells, and exhibit epithelial cell-intrinsic cytokine production, inflammatory bowel phenotype and early mortality.
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 Rabllafl allele.
The Rabllafl 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 Name||targeted mutation 1.1, Nan Gao|
|Allele Type||Targeted (Conditional ready (e.g. floxed))|
|Gene Symbol and Name||Rab11a, RAB11A, member RAS oncogene family|
|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.|
When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). Alternatively, homozygous mice may be bred together.
When using the Rab11afl mouse strain in a publication, please cite the originating article(s) and include JAX stock #024110 in your Materials and Methods section.