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Cambridge, MA, United States

Voges M.J.,Wyss Institute for Biologically Inspired Engineering | Voges M.J.,Harvard University | Voges M.J.,Chimerion Biotechnology Inc | Voges M.J.,Technical University of Delft | And 9 more authors.
Journal of Biological Engineering

Background: Plant bioengineers require simple genetic devices for predictable localization of heterologous proteins to multiple subcellular compartments.Results: We designed novel hybrid signal sequences for multiple-compartment localization and characterize their function when fused to GFP in Nicotiana benthamiana leaf tissue. TriTag-1 and TriTag-2 use alternative splicing to generate differentially localized GFP isoforms, localizing it to the chloroplasts, peroxisomes and cytosol. TriTag-1 shows a bias for targeting the chloroplast envelope while TriTag-2 preferentially targets the peroxisomes. TriTag-3 embeds a conserved peroxisomal targeting signal within a chloroplast transit peptide, directing GFP to the chloroplasts and peroxisomes.Conclusions: Our novel signal sequences can reduce the number of cloning steps and the amount of genetic material required to target a heterologous protein to multiple locations in plant cells. This work harnesses alternative splicing and signal embedding for engineering plants to express multi-functional proteins from single genetic constructs. © 2013 Voges et al.; licensee BioMed Central Ltd. Source

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