Voges, Mathias JSilver, PamelaWay, JeffreyMattozzi, Matthew2014-03-112013Voges, Mathias J, Pamela A Silver, Jeffrey C Way, and Matthew D Mattozzi. 2013. “Targeting a heterologous protein to multiple plant organelles via rationally designed 5′ mRNA tags.” Journal of Biological Engineering 7 (1): 20. doi:10.1186/1754-1611-7-20. http://dx.doi.org/10.1186/1754-1611-7-20.1754-1611http://nrs.harvard.edu/urn-3:HUL.InstRepos:11879446Background: 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.en-USProtein targetingAlternative splicingSignal embeddingLocalizationPeroxisomeChloroplastCytosolOrganelleJournal Article2014-03-1110.1186/1754-1611-7-20