Characterization of immunoglobulin heavy chain knockout rats

Eur J Immunol. 2010 Oct;40(10):2932-41. doi: 10.1002/eji.201040939.

Abstract

The rat is a species frequently used in immunological studies but, until now, there were no models with introduced gene-specific mutations. In a recent study, we described for the first time the generation of novel rat lines with targeted mutations using zinc-finger nucleases. In this study, we compare immune development in two Ig heavy-chain KO lines; one with truncated Cμ and a new line with removed JH segments. Rats homozygous for IgM mutation generate truncated Cμ mRNA with a de novo stop codon and no Cγ mRNA. JH-deletion rats showed undetectable mRNA for all H-chain transcripts. No serum IgM, IgG, IgA and IgE were detected in these rat lines. In both lines, lymphoid B-cell numbers were reduced >95% versus WT animals. In rats homozygous for IgM mutation, no Ab-mediated hyperacute allograft rejection was encountered. Similarities in B-cell differentiation seen in Ig KO rats and ES cell-derived Ig KO mice are discussed. These Ig and B-cell-deficient rats obtained using zinc-finger nucleases-technology should be useful as biomedical research models and a powerful platform for transgenic animals expressing a human Ab repertoire.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology*
  • Cell Differentiation / immunology
  • Embryonic Stem Cells / immunology
  • Graft Survival / immunology
  • Heart Transplantation / immunology*
  • Immunoglobulin Constant Regions / genetics
  • Immunoglobulin Constant Regions / immunology*
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology*
  • Immunoglobulin Isotypes / blood
  • Immunoglobulin Joining Region / genetics
  • Immunoglobulin Joining Region / immunology*
  • Lymphoid Tissue / immunology
  • Molecular Sequence Data
  • RNA / chemistry
  • RNA / genetics
  • Rats
  • Rats, Inbred Lew
  • Rats, Mutant Strains
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction
  • Specific Pathogen-Free Organisms
  • Zinc Fingers / genetics

Substances

  • Immunoglobulin Constant Regions
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Isotypes
  • Immunoglobulin Joining Region
  • RNA