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KeyGene reference: P182


The invention relates to genome editing of plant protoplasts using a complex of a CRISPR-CAS protein and an in vitro transcribed guide RNA. For stability, proteins are normally stored in buffers comprising glycerol. The inventors discovered that the removal of glycerol prior to PEG-based transfection enhances the survival of edited protoplasts. The invention thus provides an optimal protocol for genome editing protoplasts by a CRIPSR-ribonucleoprotein complex using a transfection medium that is substantially free of glycerol.


Genome editing technologies are rapidly emerging. CRISPR-system nucleases have proven to be highly agile and easy applicable. The patented technology ensures optimal survival of CRISPR-ribonucleoprotein transfected protoplasts, thereby increasing the chance of regenerating plants from edited protoplasts. As the method avoids introduction of foreign DNA, the present method is not only highly suited for gene validation, but also the protocol of choice for product development.


Efficient targeted genome editing technologies are important research tools in gene validation. Understanding the function of genes is essential for efficient breeding of improved crops. By improving the survival of edited protoplast, the present invention contributes thereto. In addition, the protocol of the invention allows for the efficient development of new traits in product development.


Click on the link for an overview of the patent family in the European Patent Register: WO2017/222370

Please note that the European Patent Register may not be extensive and/or accurate, for which KeyGene is not responsible. Please contact your patent expert for further information.

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