GILT knock-out (KO) mice contain a neomycin cassette designed to replace exons 2-7 of the Ifi30 (interferon gamma inducible protein 30 or GILT) gene. Homozygous mice exhibit defective processing and presentation of protein antigens containing disulfide bonds affecting major histocompatibility complex class-II-restricted CD4(+) T-cell responses. This strain may be useful for the study of T cell development, T cell activation, autoimmunity, cancer and infection.
Peter Cresswell, Yale School of Medicine
Ifi30 (interferon gamma inducible protein 30 or GILT) encodes a gamma-interferon inducible lysosomal thiol reductase that functions to cleave protein disulfide bonds critical for MHC class II-restricted antigen processing, cross-presentation on MHC class I, and bacterial pathogenesis. GILT is expressed constitutively in antigen processing cells, but requires gamma-interferon induction in other cells. GILT knock-out (KO) mice contain a neomycin cassette designed to replace exons 2-7 of the Ifi30 gene. Mice homozygous for the mutation are viable and fertile. GILT KO mice immunized with hen egg lysozyme (HEL) or ribonuclease A (both containing multiple disulfide bonds) exhibit one-tenth the antigen response in the proliferation recall assay. Splenic antigen presenting cells (APCs) from GILT-/- mice placed in in vitro assays with a HEL-specific T cell hybridoma (BO4) that recognizes a HEL peptide containing two cysteines fail to recognize peptide epitope. This strain may be useful for the study of antigen processing and presentation, MHC class-II-restricted CD4(+) T-cell responses, T cell development, T cell activation, autoimmunity, cancer and infection.
A targeting vector containing a neomycin resistance cassette was used to replace exons 2 through 7 of the Ifi30 (or GILT) gene. The construct was electroporated into 129S1/Sv-derived W9.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were crossed to C57BL/6 females, and offspring were backcrossed to C57BL/6J for at least 10 generations. Genotype was confirmed by Southern blot. Western blot analysis in lymph node, spleen and lung tissue demonstrates the absence of protein. Upon arrival, mice were bred to C57BL/6J for at least one generation to establish the colony.
|Allele Name||targeted mutation 1, Peter Cresswell|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Ifi30, interferon gamma inducible protein 30|
|Strain of Origin||129S1/Sv-Oca2+ Tyr+ Kitl+|
|Molecular Note||Exons 2 though 7 were replaced with a neomycin resistance gene. Western analysis demonstrated a lack of protein in homozygous mutant animals in extracts from the lymph nodes, spleen, lung and muscle. Weak signals were detected in extracts from the kidney and liver.|
When maintaining a live colony, these mice are bred by homozygote sibling matings.
When using the GILT- mouse strain in a publication, please cite the originating article(s) and include JAX stock #035475 in your Materials and Methods section.