Recombination Can Lead to Spurious Results in Retroviral Transduction with Dually Fluorescent Reporter Genes
ABSTRACT
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.
FOOTNOTES
- Received 30 August 2013.
- Accepted 20 September 2013.
- Address correspondence to Marc C. Johnson, marcjohnson{at}health.missouri.edu.
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D.J.S., D.A.E., and M.F.B. contributed equally to this article.
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Published ahead of print 25 September 2013
- Copyright © 2013, American Society for Microbiology. All Rights Reserved.
