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Freedom to operate analysis, design and evaluation of expression cassettes that confer tolerance to glyphosate
Análisis de libertad de operación, diseño y evaluación de casetes de expresión que confieren tolerancia a glifosato
DOI:
https://doi.org/10.15446/agron.colomb.v38n2.79150Keywords:
herbicide tolerance, codon usage, gene design (en)tolerancia a herbicidas, uso codónico, diseño de genes (es)
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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.
La tolerancia al herbicida glifosato es la característica predominante en los eventos transgénicos liberados comercialmente en el mundo, siendo un ejemplo del uso exitoso que ha tenido la transgénesis para mejorar el control de malezas en diferentes cultivos. Los genotipos tolerantes al glifosato han sido desarrollados por compañías multinacionales usando tecnologías protegidas por patentes. Para algunos de estos eventos todas las patentes asociadas finalizaron recientemente, y por lo tanto se han convertido en candidatos para desarrollar agrobiogenéricos resistentes a herbicidas derivados de genotipos locales a partir de investigaciones nacionales. Como primer paso del proceso, presentamos el diseño in silico de tres casetes de expresión con el fin de utilizarlos en la transformación genética de variedades colombianas de soya (Glycine max) para conferirles tolerancia al glifosato. Se transformó genéticamente la planta modelo Nicotiana benthamiana para validar la funcionalidad de los casetes y se detectó la expresión del transgén mediante RT-PCR. Adicionalmente un estudio de libertad de operación de las secuencias utilizadas en los casetes sugiere que su uso comercial en Colombia no viola derechos de terceros. Este análisis debe ser actualizado y validado por expertos en propiedad intelectual antes de la comercialización.
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1. Julian Mora-Oberlaender, Jenny Jiménez-Barreto, Yadira Rodríguez-Abril, Meike Estrada-Arteaga, Alejandro Chaparro-Giraldo. (2022). Cisgenic Crops: Potential and Prospects. Concepts and Strategies in Plant Sciences. , p.89. https://doi.org/10.1007/978-3-031-06628-3_6.
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