C;129S4-Rag2^tm1.1Flv Thpo^{tm1.1(TPO)Flv} Il2rg^tm1.1Flv/J

Stock number: 014594
Product name: Rag2- Il2rg- TPO^h
Research areas: Hematological Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

This immunodeficient strain carrying humanized Thpo (thrombopoietin) in addition to Rag2 and Il2rg knock-outs may be useful for transplantation studies, particularly those involving hematopoiesis.

Detailed description

This compound mutant strain carries deletions in the Rag2 and X-linked Il2rg genes in addition to a targeted replacement of mouse Thpo (thrombopoietin) coding sequences with those of the human gene.

Triple homozygotes (hemizygous/homozygous for the Il2rg mutation) lack T, B and NK cells and express human THPO in the same tissues as mouse Thpo, although at a lower concentration. Homozygous humanization of THPO leads to significantly increased levels of human cell engraftment in the bone marrow of hosts, and multilineage differentiation of hematopoietic cells is improved. An increased ratio of myelomonocyte versus lymphoid lineages is also seen. The self-renewal capacity of engrafted human stem and progenitor cells is improved.

The creation of this strain was funded by the Bill and Melinda Gates Foundation.

Development

An embryonic stem (ES) cell line carrying deletions in the Rag2 and Il2rg genes (** below) were re-targeted to place the human thrombopoietin gene (THPO) under the control of the mouse Thpo promoter. A loxP-flanked human ubiquitin promoter (hUBp)-neomycin selection cassette associated with the THPO targeting vector was excised by transient infection with a cre plasmid. This strain was maintained on a mixed BALB/c and 129S4 genetic background by the donating laboratory.

**V17 embryonic stem (ES) cells (derived from BALB/cTac x 129S4/SvJae mice) were used to sequentially delete the coding regions of the Rag2 and X chromosome-linked Il2rg genes. A loxP-flanked PGK-neomycin cassette was targeted to Rag2 and a loxP-flanked human ubiquitin promoter-driven hygromycin cassette was directed to Il2rg. Transient infection with a cre plasmid excised the floxed segments.

Allele

Symbol: Rag2^tm1.1Flv
Name: targeted mutation 1.1, Richard A Flavell
MGI accession ID: MGI:4887836
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Rag2
Marker name: recombination activating gene 2
Chromosome: 2
Strain of origin: BALB/cAnNTac x 129S4/SvJae
Molecular note: A floxed PGK-neomycin cassette replaced the coding region. Cre-mediated recombination removed the neo cassette.
General note: Originally generated in conjunction with Il2rg^tm1.1Flv.

Allele

Symbol: Il2rg^tm1.1Flv
Name: targeted mutation 1.1, Richard A Flavell
MGI accession ID: MGI:4887837
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Chromosome: X
Strain of origin: BALB/cAnNTac x 129S4/SvJae
Molecular note: A floxed human ubiquitin promoter-driven hygromycin cassette replaced the coding region. Cre-mediated recombination removed the hygro cassette.
General note: Originally generated in conjunction with Rag2^tm1.1Flv.

Allele

Symbol: Thpo^{tm1.1(TPO)Flv}
Name: targeted mutation 1.1, Richard A Flavell
MGI accession ID: MGI:4887867
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Thpo
Marker name: thrombopoietin
Chromosome: 16
Strain of origin: BALB/cAnNTac x 129S4/SvJae
Expressed genes: TPO,thyroid peroxidase,human
Molecular note: The coding region was replaced with the corresponding human sequence. Cre-mediated recombination removed the neo cassette.
General note: Originally generated in ES cells containing Il2rg^tm1.1Flv and Rag2^tm1.1Flv.