Single Targeted Exon Mutation Creates a True Congenic Mouse for Competitive Hematopoietic Stem Cell Transplantation: The C57BL/6-CD45.1(STEM) Mouse

Stem Cell Reports. 2016 Jun 14;6(6):985-992. doi: 10.1016/j.stemcr.2016.04.010. Epub 2016 May 12.

Abstract

Defining the molecular regulators of hematopoietic stem and progenitor cells (HSPCs) requires in vivo functional analyses. Competitive bone marrow transplants (BMTs) compare control and test HSPCs to demonstrate the functional role of a genetic change or chemical perturbation. Competitive BMT is enabled by antibodies that specifically recognize hematopoietic cells from congenic mouse strains due to variants of the cell surface protein CD45, designated CD45.1 and CD45.2. The current congenic competitor strain, B6.SJL-Ptprc(a) Pepc(b)/BoyJ (CD45.1), has a substantial inherent disadvantage in competition against the C57BL/6 (CD45.2) strain, confounding experimental interpretation. Despite backcrossing, the congenic interval over which the B6.SJL-Ptprc(a) Pepc(b)/BoyJ strain differs is almost 40 Mb encoding ∼300 genes. Here, we demonstrate that a single amino acid change determines the CD45.1 epitope. Further, we report on the single targeted exon mutant (STEM) mouse strain, CD45.1(STEM), which is functionally equivalent to CD45.2 cells in competitive BMT. This strain will permit the precise definition of functional roles for candidate genes using in vivo HSPC assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Epitopes / genetics*
  • Epitopes / immunology
  • Exons*
  • Gene Expression
  • Gene Targeting*
  • Hematopoietic Stem Cell Transplantation
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / immunology*
  • Leukocyte Common Antigens / genetics*
  • Leukocyte Common Antigens / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred Strains
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Point Mutation
  • Protein Isoforms / genetics
  • Protein Isoforms / immunology

Substances

  • Epitopes
  • Protein Isoforms
  • Leukocyte Common Antigens
  • Ptprc protein, mouse

Grant support