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    B-hRAGE mice plus

    C57BL/6N-Agertm2(AGER)Bcgen/Bcgen • 113157

    B-hRAGE mice plus

    Product nameB-hRAGE mice plus
    Catalog number113157
    Strain nameC57BL/6N-Agertm2(AGER)Bcgen/Bcgen
    Strain backgroundC57BL/6N
    NCBI gene ID177 (Human)
    AliasesRAGE; sRAGE; SCARJ1
    ApplicationThis product is used for pharmacodynamics and safety evaluation of drugs for various chronic degenerative diseases and neoplastic diseases.

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    • Description
    • Targeting strategy
    • Phenotypic analysis

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      发表文章

        Description
        • RAGE is expressed in more than 300 cells in addition to lung tissue, and studies have shown that tumors can increase the expression of this gene in a variety of cells.
        • Exons 1-11 including 3’UTR of the mouse RAGE gene that encodes the full-length protein was replaced by human RAGE exons 1-11 including 3’UTR in B-hRAGE mice plus.
        • Mouse RAGE protein was only detectable in wild-type C57BL/6 mice. Human RAGE protein was detectable in homozygous B-hRAGE mice plus but not in wild-type mice.
        • Mouse RAGE mRNA was only detectable in wild-type C57BL/6 mice. Human RAGE mRNA was detectable in homozygous B-hRAGE mice plus but not in wild-type mice.
        Targeting strategy

        Gene targeting strategy for B-hRAGE mice plus. Exons 1-11 including 3’UTR of the mouse RAGE gene that encodes the full-length protein was replaced by human RAGE exons 1-11 including 3’UTR in B-hRAGE mice plus.

        mRNA expression analysis

        Strain specific analysis of RAGE mRNA expression in wild-type C57BL/6 mice and B-hRAGE mice plus by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hRAGE mice plus (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human RAGE primers. Mouse RAGE mRNA was detectable only in wild-type C57BL/6 mice. Human RAGE mRNA was detectable only in homozygous B-hRAGE mice plus but not in wild-type mice.

        Strain specific analysis of RAGE expression in wild-type C57BL/6N mice and heterozygous humanized B-hRAGE mice plus by RT-qPCR. Blood, and blood RNA were isolated from wild-type C57BL/6N mice (+/+) (n=3, 9-week-old) and B-hRAGE mice plus (n=5, 9-week-old), then cDNA libraries were synthesized by reverse transcription, followed by real-time quantitative PCR with RAGE primers. The mRNA levels of RAGE was in detectable in B-hRAGE mice plus with Applied Biosystems SYBR, but not wild-type C57BL/6N mice. The mRNA levels of RAGE in lung from B-hRAGE mice plus was higher than that in blood from B-hRAGE mice plus.

        Protein expression analysis

        Western blot analysis of RAGE protein expression in homozygous B-hRAGE mice plus. Lung tissue lysates were collected from wild-type C57BL/6N mice (+/+), homozygous B-hRAGE mice plus (H/H) and B-Rage KO mice(RM), and then analyzed by western blot with species-specific anti-RAGE antibody. 40 μg total proteins were loaded for western blotting analysis. RAGE was detected in lung.

        Protein expression analysis in serum

        Strain specific RAGE expression analysis in wild-type C57BL/6N mice and heterozygous humanized B-hRAGE mice plus by ELISA. Serum was collected from wild-type C57BL/6N mice (+/+) (male, n=3, 7-week-old) and heterozygous B-hRAGE mice plus (H/+) (male, n=3, 15-week-old). Expression level of mouse and human RAGE were analyzed by ELISA (Human RAGE Quantikine ELISA Kit: R&D, DRG00). Human RAGE was exclusively detectable in heterozygous B-hRAGE mice plus(n=3). Values are expressed as mean ± SEM.

        Protein expression analysis in lung

        Strain specific RAGE expression analysis in wild-type C57BL/6N mice and heterozygous humanized B-hRAGE mice plus by ELISA. Lung homogenates were collected from wild-type C57BL/6N mice (+/+) (male, n=3, 7-week-old) and heterozygous B-hRAGE mice plus (H/+) (male, n=3, 15-week-old). Expression level of mouse and human RAGE were analyzed by ELISA (Human RAGE Quantikine ELISA Kit: R&D, DRG00). Human RAGE was exclusively detectable in heterozygous B-hRAGE mice plus(n=3). Values are expressed as mean ± SEM.

        B-hRAGE mice plus validation

        The inhibitory efficiency of the RAGE targeted nucleic acid drugs in homozygous B-hRAGE mice plus. B-hRAGE mice plus were randomly divided into 3 groups (5-6 weeks old, female). The human RAGE-targeted nucleic acid drug (provided by a client) and PBS were administered to the mice individually. The nucleic acid drug was administered in the form of PBS aqueous solution. The mice were sacrificed on day 7. (A) The schematic diagram of experimental processing. (B) The changes in RAGE mRNA expression levels in lung tissue on day 7 after administration, compared to the levels before administration. Values are expressed as mean ± SEM.

        * When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hRAGE mice plus] (Cat# 113157) was purchased from Biocytogen.