Coordinator

Axel LABEYRIE, F2F team -  This email address is being protected from spambots. You need JavaScript enabled to view it.

Currently, the genetic edition technique CRISPR-Cas9 is a unique opportunity to gain understanding on the role of genes. However, in plants its use is too often limited to species that can easily produce the embryogenic calli required for transfection by Agrobacterium bacteria carrying Cas9-encoding plasmids and guide sequences.

As an alternative to Agrobacterium-based transfection, it is possible to use methods relying on direct DNA transfer such as biolistics, but they may generate chimeras. Another option is to use protoplasts, which are produced through the enzymatic digestion of the cell wall and thus facilitate the integration of the exogenous DNA fragment. During the following steps of the protocol, the cell wall regenerates, organites are reorganized and cell division resumes, yielding clusters of undifferentiated cells that will multiply into micro-colonies and micro-calli. Once they are transferred to a suitable medium, these calli may then display embryogenic properties and lead to the production of somatic embryos that will regenerate full plantlets. However, if in most plant species transfected protoplasts production seems feasible, there are still many obstacles to plantlets regeneration. Our current work aim at circumventing this issue so that targeted gene edition may be routinely used in palms.

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Partners : PalmElit SAS, France ; I.A.G.E, France ; RegenCrop network, INRA, Versailles, France