C57BL/6N-Tnfsf15tm2(TNFSF15)Bcgen Rag2tm1Bcgen/Bcgen • 114011
Product name | B-hTL1A, Rag2 KO mice |
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Catalog number | 114011 |
Strain name | C57BL/6N-Tnfsf15tm2(TNFSF15)Bcgen Rag2tm1Bcgen/Bcgen |
Strain background | C57BL/6N |
NCBI gene ID | 326623,19374 (Mouse) |
Aliases | Tl1; Tl1a; Vegi; Tnlg1b; Rag; Rag-2 |
Gene targeting strategy for B-hTL1A, Rag2 KO mice.
The exons 1-4 of mouse Tl1a gene that encode extracellular domain were replaced by human counterparts in B-hTL1A, Rag2 KO mice. The genomic region of mouse Tl1a gene that encodes transmembrane domain and cytoplasmic portion was retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were also retained. The chimeric TL1A expression was driven by endogenous mouse Tl1a promoter, while mouse Tl1a gene transcription and translation will be disrupted.
The exon 3 and 3’UTR region of mouse Rag2 were knocked out in B-hTL1A, Rag2 KO mice, resulting in a disruption of the Rag2 gene.
Soluble TL1A expression analysis in B-hTL1A, Rag2 KO mice by ELISA. Bone marrow derived dendritic cells (BMDCs) were produced by culturing the bone marrow from wild-type C57BL/6N mice (+/+) , homozygous B-hTL1A, Rag2 KO mice (H/H;-/-), which were stimulated with LPS in vitro for 24 hrs. After stimulation, the supernatants were collected and the levels of soluble TL1A were measured using a species-specific human TL1A ELISA kit (R&D, DY1319-05). Soluble human TL1A was exclusively detectable in homozygous B-hTL1A, Rag2 KO mice but not wild-type C57BL/6N mice. Values are expressed as mean ± SEM. ND: not detectable.
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6N mice (female, n=3, 6-week-old), B-hTL1A mice (female, n=3, 6-week-old), B-Rag2 KO mice (female, n=3, 6-week-old) and B-hTL1A, Rag2 KO mice (female, n=3, 6-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A mice were similar to those in C57BL/6N mice. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A, Rag2 KO mice were similar to those in B-Rag2 KO mice. Values are expressed as mean ± SEM.
Frequency of leukocyte subpopulations in spleen by flow cytometry. Blood was isolated from wild-type C57BL/6N mice (female, n=3, 6-week-old), B-hTL1A mice (female, n=3, 6-week-old), B-Rag2 KO mice (female, n=3, 6-week-old) and B-hTL1A, Rag2 KO mice (female, n=3, 6-week-old). A. Flow cytometry analysis of the blood was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A mice were similar to those in C57BL/6N mice. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A, Rag2 KO mice were similar to those in B-Rag2 KO mice. Values are expressed as mean ± SEM.