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Toward transgene-free genome editing in poplar plants using Agrobacterium-mediated delivery of a CRISPR/Cas9 cytidine base editor

Abstract : We present here the first evidence of the precise targeting of point mutations in the genome of a forest tree species using a cytidine base editor (CBE). This was done using the classical Agrobacterium cocultivation method routinely used on our model hybrid poplar clone (INRA 717-1B4) for more than 30 years. Our ultimate goal is to produce transgene-free edited poplar plants. Indeed, in perennial species with long generation time, such as trees, it is virtually impossible to get rid of alien copies introduced into the plant genome during the cocultivation step. Therefore, using a strategy already shown to be successful in tomato and potato (Veillet et al., 2019), we targeted the endogenous poplar acetolactate synthase (ALS) gene by a CBE through Agrobacterium tumefaciens cocultivation. Using an optimized procedure, we were able to regenerate at high yield chlorsulfuron-resistant plants. Interestingly, a small number of these herbicide-resistant plants do not show evidence of T-DNA integration. Molecular analyses are under way to more accurately characterize these plants. Our most recent experiments aim to evaluate on this poplar model system the co-edition of ALS with another gene.
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https://hal.inrae.fr/hal-03289194
Contributor : Annabelle Déjardin Connect in order to contact the contributor
Submitted on : Friday, July 16, 2021 - 5:09:46 PM
Last modification on : Wednesday, July 21, 2021 - 3:50:03 AM

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  • HAL Id : hal-03289194, version 1

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Gilles Pilate, Florian Veillet, Orlane Touzet, Annabelle Déjardin. Toward transgene-free genome editing in poplar plants using Agrobacterium-mediated delivery of a CRISPR/Cas9 cytidine base editor. 10th annual meeting of the EFOR network, May 2021, Paris, France. ⟨hal-03289194⟩

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