Lentivirally generated eGFP-transgenic rats allow efficient cell tracking in vivo.
Here we describe the efficient generation of eGFP-transgenic rats using a lentiviral approach. Analysis of the founder generation demonstrated that 46% of the offspring had stably integrated the provirus into the genome and of those 92% expressed eGFP in all blood-derived leukocytes. In contrast to their offspring, all founder rats were mosaic with regard to eGFP-expression, suggesting delayed viral transduction after injection. The expression level of eGFP in the F1 generation is influenced by and segregates with the site of proviral integration. Interestingly, a single copy of the transgene is sufficient for reliable detection by flow cytometry, irrespective of the leukocyte subtype analyzed. Adoptive transfer of purified CD4(+) T-lymphocytes from transgenic rats and subsequent reisolation from various organs further demonstrated that expression of the lentiviral transgene is maintained in a foreign host and therefore allows for efficient tracking of transferred cells. Taken together, lentivirally generated eGFP-transgenic rats are a powerful tool for various applications in immunology and presumably also many other fields.
van den Brandt
Jens
J
Department of Molecular Immunology, Institute of Virology and Immunobiology, University of Wuerzburg, Würzburg, Germany.
Wang
Dapeng
D
Kwon
Soon-Hwan
SH
Heinkelein
Martin
M
Reichardt
Holger M
HM
D019264:Adoptive Transfer; D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics*; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic; D019076:Transgenes / Q000502:physiology
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins