Recombination can lead to spurious results in retroviral transduction with dually fluorescent reporter genes.
Fluorescent proteins are routinely employed as reporters in retroviral vectors. Here, we demonstrate that transduction with retroviral vectors carrying a tandem-dimer Tomato (TdTom) reporter produces two distinct fluorescent cell populations following template jumping due to a single nucleotide polymorphism between the first and second Tomato genes. Template jumping also occurs between repeated sequences in the Tomato and green fluorescent protein (GFP) genes. Thus, proper interpretation of the fluorescence intensity of transduced cells requires caution.
Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, Missouri, USA.
Daniel J
DJ
Salamango
David A
DA
Evans
Mariju F
MF
Baluyot
Jackson N
JN
Furlong
Marc C
MC
Johnson
D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D016662:Virus Integration
D008164:Luminescent Proteins