These Smart13 knock-in mice express a modified human CD4 marker from the mouse Il13 gene locus. They are suitable for use in applications where tracking Il13-expressing hematopoietic cells is desired.
Richard M. Locksley, Univ of California San Francisco-HHMI
The Smart13 or Il13Smart allele has a bi-cistronic IRES::CD4*F43I cassette inserted between the translational stop codon and the 3' UTR of the interleukin 13 (Il13) gene. These mice express endogenous Il13 and a cell surface membrane-anchored, non-signaling human CD4 marker. Homozygous Smart13 mice are viable and fertile. Expression of the human CD4 marker is observed in Il13-expressing cells including innate type 2 helper cells (Ih2 cells) and type 2 helper T cells (Th2). These Ih2 cells are lineage-negative innate cells that arise during type 2 immunity or in response to the epithelial cytokines IL-25 and IL-33. Ih2 cells are widely distributed in mouse tissues and are particularly prevalent in mesenteric lymph nodes, spleen, and liver.
A targeting vector was designed to insert a neomycin (neo) resistance cassette, and an internal ribosome entry site (IRES), followed by truncated human CD4 cDNA, between the stop codon and the 3' untranslated region of the interleukin 13 (Il13) gene. The truncated CD4 fragment lacking intracellular signaling capacity, also contained a point mutation resulting in an I to F substitution at position 43 of the protein, rendering the human CD4 marker incapable of interacting with mouse major histocompatibility complex class II (MHCII). The construct was electroporated into 129S4/SvJae-derived PrmCre embryonic stem (ES) cells. These PrmCre (or PC3) ES cells contain the Tg(Prm-cre)70Og transgene with the protamine promoter directing cre expression to the male germline. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females to generate a colony of Il13Smart mice lacking the neo cassette. These mice were backcrossed to BALB/cJ mice (Stock No. 000651) for at least 10 generations. They were then backcrossed with CBy.SJL(B6)-Ptprca/J (Stock No. 006584) to generate Smart13 Ly5.1/5.1 homozygotes. Upon arrival at The Jackson Laboratory, mice were bred to BALB/cJ inbred mice for at least one generation to establish a colony.
|Expressed Gene||CD4, CD4 molecule, human|
|Site of Expression|
|Allele Name||targeted mutation 2.1, Richard M Locksley|
|Allele Type||Targeted (Reporter)|
|Allele Synonym(s)||Il13Smart; IL-13Smart; Il13tm2.1(CD4)Lky; Smart13|
|Gene Symbol and Name||Il13, interleukin 13|
|Expressed Gene||CD4, CD4 molecule, human|
|Strain of Origin||129S4/SvJae-Tg(Prm-cre)70Og|
|Molecular Note||A targeting vector was designed to insert a neomycin (neo) resistance cassette, and an internal ribosome entry site (IRES), followed by truncated human CD4 cDNA, between the stop codon and the 3' untranslated region. The truncated CD4 fragment(424 aa vs. 458 aa in full-length protein) lacking intracellular signaling capacity (aa 419-424 as cytoplasmic tail in truncated form vs. aa 419-458 in full-length hCD4), and also contains a point mutation resulting in an I(ATC codon) to F(TTC codon) substitution at position 43 of the protein, rendering the human CD4 marker incapable of interacting with mouse major histocompatibility complex class II (MHCII) and useful as a marker of cells expressing IL13. Cre-mediated recombination removed the neo cassette.|
When maintaining a live colony, homozygous mice may be bred together.
When using the C.129S4(Cg)-Il13tm2.1Lky/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #018869 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.