{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T03:52:59Z","timestamp":1780631579193,"version":"3.54.1"},"reference-count":69,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T00:00:00Z","timestamp":1734652800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"International Institute of Tropical Agriculture (IITA)","award":["OPP1093845"],"award-info":[{"award-number":["OPP1093845"]}]},{"name":"The Bill and Melinda Gates Foundation\u2014Accelerated Breeding of Better Bananas","award":["OPP1093845"],"award-info":[{"award-number":["OPP1093845"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>Bananas and plantains are important staple food crops affected by biotic and abiotic stresses. The gene editing technique via Clustered Regularly Interspaced Short Palindromic Repeats associated with the Cas protein (CRISPR\/Cas) has been used as an important tool for development of cultivars with high tolerance to stresses. This study sought to develop a protocol for the construction of vectors for gene knockout. Here we use the phytoene desaturase (PDS) gene as a case study in Prata-An\u00e3 banana by the nonhomologous end junction (NHEJ) method. PDS is a key gene in the carotenoid production pathway in plants and its knockout leads to easily visualized phenotypes such as dwarfism and albinism in plants. Agrobacterium-mediated transformation delivered CRISPR\/Cas9 constructs containing gRNAs were inserted into embryogenic cell suspension cultures. This is the first study to provide an effective method\/protocol for constructing gene knockout vectors, demonstrating gene editing potential in a Brazilian banana variety. The constitutive (CaMV 35S) and root-specific vectors were successfully assembled and confirmed in transformed Agrobacterium by DNA extraction and PCR. The specificity of transformation protocols makes it possible to use the CRISPR-Cas9 technique to develop Prata-An\u00e3 banana plants with enhanced tolerance\/resistance to major biotic and abiotic factors.<\/jats:p>","DOI":"10.3390\/cimb46120865","type":"journal-article","created":{"date-parts":[[2024,12,20]],"date-time":"2024-12-20T07:42:18Z","timestamp":1734680538000},"page":"14422-14437","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Construction and Validation of CRISPR\/Cas Vectors for Editing the PDS Gene in Banana (Musa spp.)"],"prefix":"10.3390","volume":"46","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8468-5099","authenticated-orcid":false,"given":"Marcelly Santana","family":"Mascarenhas","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Feira de Santana State University, Feira de Santana 44036-900, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3033-1604","authenticated-orcid":false,"given":"Fernanda dos Santos","family":"Nascimento","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5261-7016","authenticated-orcid":false,"given":"Luana Maria Pacheco","family":"Schittino","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Federal University of Vi\u00e7osa, Vi\u00e7osa 36507-900, MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2535-2483","authenticated-orcid":false,"given":"Livia Batista","family":"Galinari","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Federal University of Vi\u00e7osa, Vi\u00e7osa 36507-900, MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6400-8034","authenticated-orcid":false,"given":"Lucymeire Souza Morais","family":"Lino","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3677-4855","authenticated-orcid":false,"given":"Andresa Priscila de Souza","family":"Ramos","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1980-9945","authenticated-orcid":false,"given":"Leandro Eugenio Cardamone","family":"Diniz","sequence":"additional","affiliation":[{"name":"Embrapa Soja, Rodovia Carlos Jo\u00e3o Strass, Londrina 86085-981, PR, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiago Ant\u00f4nio de Oliveira","family":"Mendes","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Federal University of Vi\u00e7osa, Vi\u00e7osa 36507-900, MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5476-2236","authenticated-orcid":false,"given":"Claudia Fortes","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3793-1792","authenticated-orcid":false,"given":"Janay Almeida dos","family":"Santos-Serejo","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9086-7385","authenticated-orcid":false,"given":"Edson Perito","family":"Amorim","sequence":"additional","affiliation":[{"name":"Embrapa Mandioca e Fruticultura, Cruz das Almas 44380-000, BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,20]]},"reference":[{"key":"ref_1","unstructured":"FAOSTAT (2024, February 06). 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