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Published on: April 18, 2014
Targeting a heterologous protein to multiple plant organelles via rationally designed 5' mRNA tags
Mathias J Voges1, Pamela A Silver, Jeffrey C Way
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA. matt.mattozzi@wyss.harvard.edu.
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.
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