Examinando por Materia "CRISPR/Cas9"

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    Desarrollo de líneas de cacao (Theobroma cacao L.) editadas genéticamente que limiten la acumulación de cadmio en sus tejidos : FASE-I
    (Universidad EAFIT, 2020) Rodríguez Bolaño, Jose Ángel; Marín Vélez, Neyder Alejandro; Villanueva Mejía, Diego Fernando
    Cocoa (Theobroma cacao) is the primary source of raw material for chocolate production and its derivatives. In current decades, high concentrations of cadmium have been found in cocoa-based products, which has been recognized by several organizations as a threat to human health, food security, and the future of the cocoa industry. Therefore, advances in agronomic and genetic methods that reduce this heavy metal in plants are necessary. In the present study, an in silico metadata analysis was carried out to identify genes involved in cadmium absorption to propose them as a target for genetic improvement through modern precision biotechnology techniques as genetic editing. We found 26 proteins, 88% of them reported in Arabidopsis thaliana, and 38% in Oryza sativa, which belong to 8 protein families. The remaining members of each family were included for analysis. Thus, expression data obtained from the Genevestigator software were evaluated for 44 genes of A. thaliana and 37 of O. sativa, and it was found that the TcNRAMP5 gene of T. cacao, "orthologs" of OsNRAMP5 of O. sativa, is closely related to the root absorption process of Cd. Consequently, it was postulated as a gene target for genetic editing to decrease Cd concentration in cocoa plants. A genetic editing methodology based on CRISPR/Cas9 system was structured using Agrobacterium tumefaciens mediated genetic transformation and protoplast transfection. Furthermore, this research structured a methodology for the establishment and generation of in vitro plants through indirect somatic embryogenesis and protoplast isolation, as a fundamental basis of the proposed modern biotechnological platform, which leads to the improvement of genetic accuracy in cocoa.
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