The 2A-like sequences from members of the picornavirus family were utilized to construct a tricistronic vector bearing the human iduronidase (IDUA) gene along with the firefly luciferase and DsRed2 reporter genes. The 2A-like sequences mediate a cotranslational cleavage event resulting in the release of each individual protein product. Efficient cleavage was observed and all three proteins were functional in vitro and in vivo, allowing for supratherapeutic IDUA enzyme levels and the coexpression of luciferase and DsRed2 expression, which enabled us to track gene expression.
Bibliographical noteFunding Information:
The authors express their gratitude to Kevin Tram and Andrew Price for their technical assistance and advise. This work was supported in part by a University of Minnesota Children's Cancer Research Fund award (MJO) and NIH grants R01 HL49997 and HL52592 (BRB).
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