An inducible Tet-Off-H2B-GFP lentiviral reporter vector for detection and in vivo isolation of label-retaining cells

Exp Cell Res. 2010 Jul 1;316(11):1885-95. doi: 10.1016/j.yexcr.2010.02.015. Epub 2010 Feb 18.


Many regenerative cells are label-retaining cells (LRCs) due to their ability to keep a DNA label over a prolonged time. Until recently, isolation of vital LRCs was hampered due to the necessary use of fixation methods. To circumvent this, we generated a lentiviral-(HIV-1) based vector expressing a Tet-Off controlled histone 2B-GFP (Tet-Off-H2B-GFP) reporter gene for the detection and isolation of viable LRCs. In initial experiments, the vector was successfully used to infect 2- and 3-dimensional tissue culture models. Infected cultures from skin and pancreatic cells showed a very tight regulation of H2B-GFP, were sensitive to minimal amounts of doxycycline (Dox) and had a stable transgenic expression over the time of this study. Our lentiviral vector represents a reliable and easy to handle system for the successful infection, detection and isolation of LRCs from various tissues in vitro, in vivo and ex vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Cycle
  • Cell Line
  • Doxycycline / pharmacology
  • Gene Expression / drug effects
  • Genes, Reporter*
  • Genetic Engineering
  • Genetic Vectors*
  • Green Fluorescent Proteins / genetics
  • Histones / genetics*
  • Humans
  • Lentivirus / genetics*
  • Mice
  • Recombinant Fusion Proteins / genetics
  • Transfection


  • Histones
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Doxycycline