| Texto completo | |
| Autor(es): Mostrar menos - |
Rocha, Dhiovanna Correia
;
Omoregbee, Miracle Osazee
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Contiliani, Danyel Fernandes
;
Mandlik, Rushil
;
Li, Gen
;
Mascoveto, Juliet
;
Coleman, Gary
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Culver, James N.
;
Leal, Daniel Rodriguez
;
de Souza, Alessandra Alves
;
Qi, Yiping
Número total de Autores: 11
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| Tipo de documento: | Artigo Científico |
| Fonte: | Plant Cell Reports; v. 44, n. 11, p. 17-pg., 2025-10-22. |
| Resumo | |
Key messageTransgene-free genome editing of the gene of interest in citrus and poplar has been achieved by co-editing the ALS gene via transient transgene expression of an efficient cytosine base editor.AbstractCRISPR-Cas genome editing systems have been widely used in plants. However, such genome-edited plants are nearly always transgenic in the first generation when Agrobacterium-mediated transformation is used. Transgene-free genome-edited plants are valuable for genetic analysis and breeding as well as simplifying regulatory approval. It can be challenging to generate transgene-free genome-edited plants in vegetatively propagated or perennial plants. To advance transgene-free genome editing in citrus and poplar, we investigated a co-editing strategy using an efficient cytosine base editor (CBE) to edit the ALS gene to confer herbicide resistance combined with transient transgene expression and potential mobile RNA-based movement of CBE transcripts to neighboring, non-transgenic cells. An FCY-UPP based cytotoxin system was used to select non-transgenic plants that survive after culturing on 5-FC containing medium. While the editing efficiency is higher in poplar than in citrus, our results show that the CBE-based co-editing strategy works in both citrus and poplar, albeit with low efficiency for biallelic edits. Unexpectedly, the addition of the TLS mobile RNA sequence reduced genome editing efficiency in both transgenic and non-transgenic plants. Although a small fraction of escaping plants is detected in both positive and negative selection processes, our data demonstrate a promising approach for generating transgene-free base-edited plants. (AU) | |
| Processo FAPESP: | 21/03466-7 - Edição de genes de Citrus sinensis visando atenuar o acúmulo de caloses associadas aos sintomas do Huanglongbing (HLB) |
| Beneficiário: | Dhiôvanna Corrêia Rocha |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 20/07045-3 - Estratégias biotecnológicas e genômicas para qualidade, produtividade e manejo sustentável de citros, café e cana-de-açúcar no estado de São Paulo |
| Beneficiário: | Mariângela Cristofani-Yaly |
| Modalidade de apoio: | Auxílio à Pesquisa - Núcleos de Pesquisa Orientada a Problemas em São Paulo |
| Processo FAPESP: | 23/09068-9 - Novas tecnologias para a transfecção do complexo RNP e obtenção de variedades comerciais de Citrus sinensis editadas DNA-free |
| Beneficiário: | Dhiôvanna Corrêia Rocha |
| Modalidade de apoio: | Bolsas no Exterior - Estágio de Pesquisa - Doutorado |