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Generation of a plasma cell-specific inducible Cre transgenic mouse

浆细胞特异性诱导型 Cre 转基因小鼠的产生

基本信息

批准号：
10375383
负责人：
Joel R Wilmore
金额：
$ 8.1万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-19 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10375383
关键词：
Alleles Antibodies Antibody-Producing Cells Antigens Autoantibodies Autoimmune Diseases Autoimmunity B cell differentiation B-Cell Antigen Receptor B-Lymphocytes Bacterial Artificial Chromosomes Bone Marrow C57BL/6 Mouse CD19 gene CRISPR/Cas technology Cell Differentiation process Cell Lineage Cell Maintenance Cell Maturation Cell Survival Cell physiology Cells Cellular biology Critical Pathways Disease Embryonic Development Ensure Enterobacteria phage P1 Cre recombinase Flow Cytometry Gene Deletion Gene Expression Generations Genes Genetic Genetic Transcription Goals Humoral Immunities Immunoglobulin A Immunoglobulin G Immunoglobulin-Secreting Cells Immunologic Memory Immunologist In Vitro Infection Investigation Knock-in Label Lead Longevity MCL1 gene Maintenance Malignant Neoplasms Modeling Multiple Myeloma Mus Organ Transplantation Pathogenicity Phenotype Plasma Cells Play Population Research Role Site Source Specificity Spleen Structure of germinal center of lymph node TNFRSF17 gene Tamoxifen Techniques Technology Terminator Codon Testing Transgenes Transgenic Mice Transposase Untranslated RNA Vaccination Vaccine Design Vaccines Validation Work cytokine genetic manipulation immune function improved mouse model mucosal site novel novel strategies promoter protein B response tool transcription factor vaccine efficacy

项目摘要

Project Summary/Abstract Plasma cells produce antibodies in response to infection or vaccination, but also play pathogenic roles in autoimmunity and organ transplant. Despite the critical role of plasma cells in humoral immunity, there is currently no transgenic mouse model that allows for conditional deletion of genes specifically in plasma cells. This proposal aims to generate and validate a mouse model that expresses an inducible Cre specifically in plasma cells. This goal will be achieved by using piggyBac transposase to insert a BAC cassette containing the tamoxifen inducible CreERT2 gene upstream of the Tnfrsf17 locus. Tnfrsf17 encodes the protein B cell maturation antigen (BCMA) and is expressed exclusively in plasma cells. Since BCMA is critical for plasma cell survival and knock-in or CRISPR/Cas9 techniques can result in a null allele, BAC technology provides advantageous alternative because it leaves endogenous gene expression intact. Aim 1 will set out to generate the mouse model and Aim 2 will be dedicated to validating the specificity and efficiency of the model. To test specificity to the plasma cell lineage, the BCMA-CreERT2 mouse will be crossed to the lineage-tracing mouse R26R-EYFP. When these mice are treated with tamoxifen the plasma cells will activate Cre recombinase and delete a loxP flanked stop codon upstream of Rosa26 driven EYFP, irreversibly labeling the plasma cells. Since loxP sites at the Rosa26 locus are notoriously sensitive to Cre, we will also test the ability of the BCMA- CreERT2 mouse to efficiently delete the Mcl-1 gene from Mcl-1fl/fl mice. Long-lived plasma cells require Mcl-1 for survival and therefore efficient deletion will result in the loss of the standing pool of long-lived plasma cells. Upon completion of these aims, the BCMA-CreERT2 mouse will be a valuable tool for the analysis of long-lived plasma cells leading to research that will advance the treatment of a variety of diseases.

项目总结/文摘