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NIH Research Festival

September 16 – 18, 2015

Generation of CD40 mutant mice by CRISPR-Cas9 Technology

Thursday, September 17, 2015 – Poster Session III
3:30 – 5:00 p.m.

FAES Terrace

NCI

IMMUNO-28

Authors

  • LE Rivera Hiraldo
  • J Chiang
  • R Hodes

Abstract

CD40 is a member of the tumor necrosis factor receptor (TNFR) family expressed by multiple tissues including dendritic cells, B cells and thymic medullary epithelial cells. CD40 is required for proper functioning of the immune system including negative selection of self-reactive thymocytes, isotype switching of immunoglobulin (Ig) in antigen-stimulated B cells, and production of cytokines and chemokines in activated APCs. However, the cell type which must express CD40 in order to drive these various processes has yet to be clearly established. In order to investigate the cell-specific requirements for CD40, we are working to generate a collection of CD40 mutants using CRISPR-Cas9, which is a RNA guided endonuclease that can be used for targeted genome editing. Among the CD40 variants that we will generate are a conditional (cKO) and reversible (rKO) knockout. This will be accomplished using Cre-Lox recombination to remove CD40 from certain cell types (cKO) or restrict expression to specific cell types (rKO). These approaches will provide critical information on the cell-specific functions of CD40. We will also be generating mice in which we have modified the CD40 intracellular domain to probe further the CD40 signal transduction requirements for its various biological functions. Lastly, we are generating a mouse-human CD40 (mhCD40) chimeric mouse that can be used as an animal model to study functional consequences of polymorphisms in human CD40. In summary the use of the CD40 variants described above should allow elucidation of the CD40 cell-specific signaling requirements for normal immune responses.

Scientific Focus Area: Immunology

This page was last updated on Friday, March 26, 2021

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