AUTHOR=Prado Guilherme Souza , Rocha Dhiôvanna Corrêia , Santos Lucas Nascimento dos , Contiliani Danyel Fernandes , Nobile Paula Macedo , Martinati-Schenk Juliana Camargo , Padilha Lilian , Maluf Mirian Perez , Lubini Greice , Pereira Tiago Campos , Monteiro-Vitorello Claudia Barros , Creste Silvana , Boscariol-Camargo Raquel Luciana , Takita Marco Aurélio , Cristofani-Yaly Mariângela , Souza Alessandra Alves de TITLE=CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane JOURNAL=Frontiers in Plant Science VOLUME=14 YEAR=2024 URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1331258 DOI=10.3389/fpls.2023.1331258 ISSN=1664-462X ABSTRACT=

Gene editing technologies have opened up the possibility of manipulating the genome of any organism in a predicted way. CRISPR technology is the most used genome editing tool and, in agriculture, it has allowed the expansion of possibilities in plant biotechnology, such as gene knockout or knock-in, transcriptional regulation, epigenetic modification, base editing, RNA editing, prime editing, and nucleic acid probing or detection. This technology mostly depends on in vitro tissue culture and genetic transformation/transfection protocols, which sometimes become the major challenges for its application in different crops. Agrobacterium-mediated transformation, biolistics, plasmid or RNP (ribonucleoprotein) transfection of protoplasts are some of the commonly used CRISPR delivery methods, but they depend on the genotype and target gene for efficient editing. The choice of the CRISPR system (Cas9, Cas12), CRISPR mechanism (plasmid or RNP) and transfection technique (Agrobacterium spp., PEG solution, lipofection) directly impacts the transformation efficiency and/or editing rate. Besides, CRISPR/Cas technology has made countries rethink regulatory frameworks concerning genetically modified organisms and flexibilize regulatory obstacles for edited plants. Here we present an overview of the state-of-the-art of CRISPR technology applied to three important crops worldwide (citrus, coffee and sugarcane), considering the biological, methodological, and regulatory aspects of its application. In addition, we provide perspectives on recently developed CRISPR tools and promising applications for each of these crops, thus highlighting the usefulness of gene editing to develop novel cultivars.