The Wt1flox allele has loxP sites flanking exons 8-9 of the Wilms tumor 1 homolog gene. Removal of the floxed sequence creates a null allele. These mice may be useful for generating tissue-specific WT1-deletion for studying the role of this zinc finger nuclear transcription factor in leukemias and solid tumors.
Vicki Huff, University of Texas, M. D. Anderson Cancer Center
The Wilms' tumor (WT) suppressor gene (Wt1) encodes a zinc finger nuclear transcription factor. WT1 functions in cell growth and differentiation. Congenital WT1 mutations predispose to Wilms tumor (a tumor of the kidneys), and are observed in individuals with Denys-Drash Syndrome, Frasier Syndrome and WAGR (Wilms tumour, aniridia [absence of iris], genitourinary anomalies and retardation). A combination of germline and somatic mutations are observed in a subset of Wilms tumor where they are almost invariably homozygous - resulting in inactivation of WT1. Somatic WT1 mutations are present in acute and chronic myeloid leukemia, myelodysplastic syndrome and other blood neoplasms (including acute lymphoblastic leukemia). Wilms tumors display WT1 expression similar to fetal kidney, consistent with the fetal origin of Wilms tumors.
In mice, germline knockout of WT1 (Wt1-/-) results in apoptosis of intermediate mesoderm and subsequent failure of kidney and gonadal development. Wt1-/- embryos die during fetal development or at birth, depending upon strain background. Somatic mosaic inactivation of Wt1 results in the development of Wilms tumor in the background of biallelic expression of the normally imprinted Igf2 gene
The Wt1 floxed allele (Wt1flox) has loxP sites flanking Wt1 exons 8-9 (encoding the second and third of the four DNA-binding zinc-finger domains). Mice homozygous for this floxed allele (Wt1flox/flox) are viable and fertile with no reported gross physical or behavioral abnormalities. The donating investigator reports that homozygotes have normal expression levels of WT1 mRNA and protein.
When bred to mice that express Cre recombinase, the resulting offspring will have the floxed region deleted in cre-expressing tissues. The deletion allele (Wt1Δ) encodes an in-frame truncated protein that is expressed at levels similar to endogenous WT1, but the Wt1Δ protein is non-functional. Homozygous deletion (Wt1Δ/Δ) embryos are phenotypically like Wt1-/- embryos - consistent with Wt1Δ being a loss-of-function allele.
The Wt1flox allele was created in the laboratory of Dr. Vicki Huff (University of Texas, M. D. Anderson Cancer Center) to have loxP sites flanking exons 8-9. First, a targeting vector was designed to place a loxP site upstream of exon 8, and a loxP-flanked neo cassette just downstream of exon 9 of the Wilms tumor 1 homolog locus (Wt1) on chromosome 2. The construct was electroporated into 129S7/SvEvBrd-Hprt+-derived AB1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric males were bred with C57BL/6 females to establish the colony. The resulting Wt1neo-loxP mice were bred with CMV-Cre transgenic mice (unspecified genetic background), and offspring with germline deletion of the neo cassette, but retaining the loxP-flanked exons 8-9, were selected (and the cre-expressing transgene was removed). The donating investigator reports Wt1flox mice (mixed C57BL/6;129 genetic background) were bred together for several years as homozygotes prior to sending males to The Jackson Laboratory Repository in 2017. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
|Allele Name||targeted mutation 2, Vicki Huff|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||Wt1flox; Wt1fx|
|Gene Symbol and Name||Wt1, Wilms tumor 1 homolog|
|Gene Synonym(s)||Wt-1; NPHS4; D630046I19Rik; GUD; Wt-1; WT33; WAGR; AWT1; WIT-2; RIKEN cDNA D630046I19 gene; D630046I19Rik|
|Strain of Origin||129S7/SvEvBrd-Hprt<+>|
|Molecular Note||Exons 8 and 9 were flanked by loxP sites and a neo resistance cassette with a loxP site was inserted downstream of the floxed exons via homologous recombination. The neo cassette was removed by mating with CMV-Cre transgenic mice.|
Mice homozygous for this floxed allele (Wt1flox/flox) are viable and fertile with no reported gross physical or behavioral abnormalities. 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.
We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.
Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.
|Frozen Mouse Embryo||$2,595.00 per straw or vial|
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCT(S)" means biological materials supplied by JACKSON, and their derivatives. "SERVICES" means projects conducted by JACKSON for other parties that may include but are not limited to the use of MICE or PRODUCTS. "RECIPIENT" means each recipient of MICE, PRODUCTS, or SERVICES provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE, PRODUCTS or SERVICES from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON’s prior written authorization.
MICE, PRODUCTS AND SERVICES ARE PROVIDED "AS IS". JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of, PRODUCTS or SERVICES, JACKSON will, at its option, provide credit or replacement for the PRODUCT received or the SERVICES provided; JACKSON makes no other representations and this shall be the exclusive remedy of the purchaser. Please note specific policy for live mice.
Consistent with the requirement for a written understanding regarding animal care and use, the JACKSON Animal Care and Use Committee will review the animal care and use protocol(s) associated with any SERVICES to be performed at JACKSON, and JACKSON shall have ultimate responsibility and authority for the care of animals while on site or in JACKSON custody.
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS, or SERVICES, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS, or SERVICES from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE, PRODUCTS or SERVICES are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or SERVICES. In addition, special terms and conditions of sale of certain MICE, PRODUCTS, or SERVICES may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and SERVICES by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or SERVICES shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or SERVICES by JACKSON.