Expression of glial fibrillary acidic protein (Gfap can be totally ablated from the jejunum and ileum in these transgeneic mice by treatment with ganciclovir, making these mice useful in studies of inflammatory bowel disease and neurogenesis.
Christine Denny, Columbia University
Dr. Rene Hen, Columbia University
Mice hemizygous for the transgenic insert are viable, normal in size, and do not display any behavioral abnormalities. Transgenic males are infertile. Proliferating cells that express the herpes simplex virus thymidine kinase (HSV-TK) transgene will metabolize ganciclovir (GCV) to toxic nucleotide analogues and undergo cell death. Transgene-derived HSV-TK is present exclusively in cells expressing endogenous Gfap. This coexpression occurs in brain astrocytes and adult neural stem cells, enteric glia, hepatic stellate cells, and unknown cells in heart, lung, kidney, adrenal, and spleen. Chronic GCV treatment for 21 days depletes GFAP-positive adult neural stem cells from forebrain proliferative zones. GCV treatment eliminated growth of primary multipotent neurospheres cultured from the germinal zones of postnatal and adult, but not early embryonic, transgenic mice. Notably, the same treatment prevented growth of secondary multipotent neurospheres from all three developmental stages. Stab or spinal cord injury with high dose GCV treatment for 7 days induces reactive astroctye cell death leading to altered leukocyte trafficking and impaired injury healing. High dose GCV treatment for 14 days ablates GFAP-positive glia from the jejunum and ileum leading to fulminant and fatal jejuno-ileitis. This mutant can be used instudies of adult neurogenesis, nervous system injury/repair mechanisms, and inflammatory bowel disease.
A herpes simplex virus thymidine kinase (HSV-TK) construct was inserted into the first exon of a mouse glial fibrillary acidic protein (GFAP) promoter cassette (clone 445). This cassette contains all the introns, promoter regulatory elements, exons, and 2.5 kb of 5' and 2 kb of 3' flanking regions of Gfap. Transgenic Gfap expression is prevented because a small segment of exon 1 has been removed in the clone. The Gfap-HSV-Tk fusion gene was injected into the male pronucleus of fertilized eggs from superovulated female C57BL/10 x CBA mated with CFLP outbred Swiss albino males. Two-cell stage eggs were implanted into pseudopregnant foster mothers. Founder line 7.1 was obtained and backcrossed to 129S6/SvEvTac mice by the donating investigator for more than 13 generations prior to sending females and males to The Jackson Laboratory Repository.
|Expressed Gene||HSV-TK, herpes simplex virus thymidine kinase,|
|Site of Expression|
|Allele Name||transgene insertion 7.1, Michael V Sofroniew|
|Allele Type||Transgenic (Inserted expressed sequence)|
|Allele Synonym(s)||GFAP-TK; Gfap-HSV-Tk|
|Gene Symbol and Name||Tg(Gfap-TK)7.1Mvs, transgene insertion 7.1, Michael V Sofroniew|
|Gene Synonym(s)||GFAP-TK; Gfap-HSV-Tk|
|Promoter||Gfap, glial fibrillary acidic protein, mouse, laboratory|
|Expressed Gene||HSV-TK, herpes simplex virus thymidine kinase,|
|Strain of Origin||(C57BL/10 x CBA)F1 X CFLP|
|General Note||Line 7.1, estimated to carry >50 copies of the transgene, was selected from three original lines (the others unnamed) for further analysis because mice of this line exhibited the greatest HSV thymidine kinase expression in the brain. |
Ganciclovir is metabolized to toxic nucleotide analogs in cells expressing HSV-TK. Subcutaneous or intraperitoneal administration of ganciclovir (GCV) or elaidic acid ganciclovir (eGCV) to transgenic mice results in specific ablation of proliferating cells that express GFAP.
|Molecular Note||The Herpes simplex virus thymidine kinase gene, including the polyadenylation signal, was ligated into the first exon of a mouse glial fibrillary acidic protein promoter cassette that contains all the exons, introns and promoter regulatory elements and includes 2.5 kb of 5' and 2 kb of 3' flanking genomic DNA. The GFAP protein is not expressed from the transgene because a small segment of exon one has been deleted. RT-PCR analysis showed co-expression of HSV-Tk and of Gfap mRNA (from the endogenous gene) in heart, lung, liver, spleen, adrenal, kidney and GI tract, as well as in brain and trigeminal ganglion; in neural tissues the co-expressing cells have the appearance of astroglia.|
|Mutations Made By|| |
Michael Sofroniew, University of California Los Angeles
|Please inquire about possible genotypes.|
The average number of mice provided from recovery of our cryopreserved strains is 10. The total number of animals provided,
their gender and genotype will vary. We will fulfill your order by providing at least two pair of mice, at least one animal of
each pair carrying the mutation of interest. Please inquire if larger numbers of animals with specific genotype and genders
are needed. Animals typically ship between 10 and 14 weeks from the date of your order. If a second cryorecovery is needed in
order to provide the minimum number of animals, animals will ship within 25 weeks.
The genotypes of animals provided may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation, or that discussed in the strain description. Please inquire about possible genotypes which will be recovered for this specific strain. The Jackson Laboratory 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.
"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.