Tracking and Selection of Retrovirally Transduced Murine Bone Marrow Cells Using Green Fluorescent Protein

  • Jessamyn Bagley
  • John Iacomini
Part of the Methods in Molecular Biology book series (MIMB, volume 183)

Abstract

Murine replication-defective retroviral vectors are among the most wellstudied, commonly used gene transfer vehicles (1,2). The authors have used retroviruses as gene delivery tools, to genetically modify bone marrow (BM) hematopoietic progenitors, in order to induce immunological tolerance to various antigens following BM transplantation (3, 4, 5). However, one of the major problems with retroviral infection of BM is poor efficiency, resulting in the low ratio of transduced to nontransduced bone marrow cells (BMCs) used for transplantation. This limitation of retroviral gene therapy may be overcome by including, within retroviral constructs, marker genes that would allow one to obtain relatively pure populations of transduced cells by cell sorting, based on marker-gene expression prior to BM transplantation. An additional benefit is that expression of a marker gene allows one to easily monitor virus transduction in vitro, and to track the fate of transduced BMCs and their progeny in vivo.

Keywords

Phenol DMSO Titration Syringe CaCl2 

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Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Jessamyn Bagley
    • 1
  • John Iacomini
    • 1
  1. 1.Transplantation Biology Research Center and Harvard Medical SchoolBoston

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