Freedom to operate analysis, design and evaluation of expression cassettes that confer tolerance to glyphosate

Abstract

Tolerance to the herbicide glyphosate is the most extended feature in commercial transgenic events released worldwide and it is an example of the successful use of genetic modification to improve weed control in crops. Glyphosate-tolerant genotypes have been developed by multinational corporations using patent- protected technologies. For some of these events, all associated patents have expired and, therefore, have become a good target for the development of herbicide-tolerant agbiogenerics by national research institutions, using local crop varieties. As a first step in this process, we present the design (in silico) of three expression cassettes with the purpose of using them in the transformation of Colombian soybean (Glycine max) varieties to confer them tolerance to glyphosate. We transformed Nicotiana benthamiana as a model to validate the functionality of the expression cassettes and detected the expression of the transgene by RT-PCR. Additionally, a Freedom to Operate analysis of the sequences used in the expression cassettes suggests that their commercial use in Colombia does not infringe third party rights. This analysis must be updated and validated by intellectual property experts prior to commercialization.