RGB marking facilitates multicolor clonal cell tracking

Nat Med. 2011 Apr;17(4):504-9. doi: 10.1038/nm.2338. Epub 2011 Mar 27.


We simultaneously transduced cells with three lentiviral gene ontology (LeGO) vectors encoding red, green or blue fluorescent proteins. Individual cells were thereby marked by different combinations of inserted vectors, resulting in the generation of numerous mixed colors, a principle we named red-green-blue (RGB) marking. We show that lentiviral vector-mediated RGB marking remained stable after cell division, thus facilitating the analysis of clonal cell fates in vitro and in vivo. Particularly, we provide evidence that RGB marking allows assessment of clonality after regeneration of injured livers by transplanted primary hepatocytes. We also used RGB vectors to mark hematopoietic stem/progenitor cells that generated colored spleen colonies. Finally, based on limiting-dilution and serial transplantation assays with tumor cells, we found that clonal tumor cells retained their specific color-code over extensive periods of time. We conclude that RGB marking represents a useful tool for cell clonality studies in tissue regeneration and pathology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Tracking / methods*
  • Clone Cells / cytology*
  • Clone Cells / metabolism*
  • Color
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hepatocytes / cytology
  • Hepatocytes / metabolism
  • Liver Regeneration
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred NOD
  • Mice, SCID
  • Mice, Transgenic
  • Neoplasm Transplantation
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transduction, Genetic
  • Tumor Cells, Cultured / metabolism
  • Tumor Cells, Cultured / pathology


  • Luminescent Proteins
  • Recombinant Proteins
  • red fluorescent protein
  • Green Fluorescent Proteins