Recombinant cystatins in plants

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Several studies documented the usefulness of recombinant protease inhibitors for the development of insect-resistant transgenic crops, but a number of recent studies also described unintended effects of these proteins modifying the host plant's metabolic characteristics. Based on novel experimental approaches involving functional proteomics and transient protein expression, this project will aim at characterizing further the effects of protease inhibitors in planta, and at identifying the enzymes targeted by these inhibitors. Genetically engineered protease inhibitor variants with different affinities for the host plant's target enzymes will then be selected to test the feasibility of avoiding, or by contrast amplifying, the unintended effects observed. From a practical viewpoint, the general goal of this project, which is not only to identify the endogenous targets of a foreign protein in planta but also to modulate its resulting effects, will represent a first 'proof-of-concept' of our ability to rationally controlling the fine balance between the intended and unintended effects of a biologically-active foreign protein expressed in genetically modified crops.