Genomic editing in Citrus L. for resistance to citrus tristeza virus
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Abstract
Context: The Citrus tristeza disease is a threat to citrus production worldwide. The search for effective strategies to develop plants resistant to the causal agent of this disease is a priority in agricultural research.
Objective: To propose a strategy to obtain plants resistant to citrus tristeza virus by genome editing in Citrus L. species.
Methods: Candidate genes were identified through a systematic review, and guide RNA sequences and gene constructs were designed for genome editing using the CRISPR/Cas system.
Results: The viral genes p20, p23 and p25 were selected as candidates for genome editing in Citrus species. Three hairpin RNA sequences with an intronic spacer for silencing and guide RNAs for insertion of these sequences were designed using the CRISPR/Cas9 system. Finally, insertion of ribointerference precursor constructs into the actin gene is proposed as a strategy to silence viral effectors critical for evading the plant defense response.
Conclusions: Genome editing is proposed as an effective strategy to control citrus tristeza disease. Following a systematic literature review, the viral genes p20, p23 and p25 were identified as targets for silencing viral effectors essential for evading plant defense. In addition, ribointerference constructs were designed for insertion into the actin gene using CRISPR/Cas9.
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