These Tgfb1flox-ex 6 mutant mice harbor loxP sites flanking exon 6 of the Tgfb1 (transforming growth factor, beta 1) gene and may be useful in generating conditional mutations for studying the role of TGFβ1 signaling in cancer, wound healing, tissue fibrosis, and muscular, neurological, ophthalmic, cardiovascular and immunological disorders, as well as other human diseases including Marfan syndrome and Camurati-Engelmann disease.
Thomas Doetschman, University of Arizona
Mice homozygous for this Tgfb1flox-ex 6 conditional allele are viable and fertile, with loxP sites flanking exon 6 of the Tgfb1 (transforming growth factor, beta 1) gene. When bred to mice that express Cre recombinase, the resulting offspring will have the floxed region deleted in the cre-expressing tissues while leaving the precursor pro-region and neighboring B9d2 (LOC232987) gene intact.
A targeting vector was designed to place a loxP-flanked PMC1-Neo cassette just upstream of exon 6, and a third loxP site just downstream of exon 6 of the targeted gene. This construct was electroporated into 129S6/SvEvTac-derived KG-1 embryonic stem (ES) cells. Correctly targeted ES cells were then transiently transfected with a Cre recombinase-expressing plasmid. The resulting ES cells with the Tgfb1flox-ex 6 genotype (neo selection cassette removed; leaving a single loxP site upstream of exon 6 and a single loxP site downstream of exon 6) were injected into recipient blastocysts. Chimeric males were bred with Black-Swiss females to generate the Tgfb1flox-ex 6 colony. Homozygous mice were bred together for approximately five generations and then bred with Tgfb1- mice (on a mixed CF-1, 129 and BALB/c genetic background; see Stock No. 002098). The Tgfb1flox-ex 6/- mice were bred together for approximately ten generations. Offspring homozygous for the Tgfb1flox-ex 6 allele were sent to The Jackson Laboratory. Upon arrival, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the Tgfb1flox-ex 6 colony.
In September 2011, the living Tgfb1flox-ex 6 mouse colony at The Jackson Laboratory Repository was found to be homozygous for the Tgfb1flox-ex 6 allele and to also be segregating for the Cre recombinase gene. The Cre recombinase expression was unlikely to be in germline (as the colony was homozygous for the Tgfb1flox-ex 6 allele using an assay that would not produce a PCR product for a deletion allele). Whether or not the source of Cre recombinase was from previously mating Tgfb1flox-ex 6 mice with LckCre mice is not known. Mice double confirmed to be Cre-negative were selected and bred to C57BL/6J inbred mice to repopulate the colony. By May 2012, the living Tgfb1flox-ex 6 mouse colony at The Jackson Laboratory Repository was homozygous for the Tgfb1flox-ex 6 mutation and without Cre recombinase.
|Allele Name||targeted mutation 2.1, Thomas Doetschman|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||Tgfb1flox ex6|
|Gene Symbol and Name||Tgfb1, transforming growth factor, beta 1|
|Gene Synonym(s)||CED; DPD1; LAP; TGF-beta 1; TGF-beta1; TGFB; TGFbeta; TGFbeta1; Tgfb; Tgfb; Tgfb-1; Tgfb-1|
|Strain of Origin||129S6/SvEvTac|
|Molecular Note||A floxed neomycin cassette was placed upstream of exon 6 with an additional loxP site placed downstream of exon 6. The neomycin cassette was subsequently removed by transient expression of cre recombinase leaving exon 6 floxed. Germline transmission of the allele was confirmed by genomic PCR.|
|Mutations Made By|| |
Thomas Doetschman, University of Arizona
When maintaining a live colony, heterozygous or homozygous mice may be bred together.
When using the Tgfb1flox ex6 mouse strain in a publication, please cite the originating article(s) and include JAX stock #010721 in your Materials and Methods section.
"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.