Percorrer por autor "Ferreira, Patrick"
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- CRISPR-CAS9 em plantas: desafios éticos, científicos e oportunidadesPublication . Choupina, Altino; Vieira, Kelly; Ferreira, PatrickA edição genómica por meio do sistema CRISPR-Ca9 representa um avanço significativo na biotecnologia, especialmente no melhoramento genético vegetal, ao permitir a modificação precisa dos genes específicos para enfrentar os desafios contemporâneos da agricultura. Este estudo tem como objetivo principal realizar uma revisão sistemática da literatura sobre o funcionamento da tecnologia CRISPR-Cas9, destacando as suas vantagens relativamente às técnicas anteriores e discutindo as suas aplicações na proteção vegetal, com ênfase no controlo de pragas, na resistência a stresse bióticos e abióticos, e no fortalecimento da imunidade vegetal. Além disso, o estudo visa refletir sobre os aspetos éticos, políticos e regulamentares envolvidos no uso dessa tecnologia. O trabalho está organizado da seguinte forma: Na Introdução apresenta-se a metodologia, objetivos e estrutura do presente estudo. No Capítulo I, apresentam-se os fundamentos do sistema CRISPR-Cas9, abordando a sua história, mecanismos de ação, além das suas aplicações médicas, farmacêuticas e agrícolas. Em seguida, no Capítulo II, explora-se o uso dessa tecnologia na proteção de plantas, analisando como as ferramentas moleculares de CRISPR-Cas9 são empregues para aumentar a resistência vegetal contra o stresse abióticos e bióticos. No Capítulo III, aprofunda-se a edição genómica de parasitas como insetos, fungos e oomicetes, discutindo o seu papel no controlo sustentável de pragas agrícolas. No Capítulo IV , introduz-se a técnica de RNA de interferência (RNAi), uma abordagem alternativa ao CRISPR-Cas9, destacando os seus avanços para conferir imunidade vegetal e resistência a stresse bióticos, além de comparar ambas as técnicas em termos de eficácia e aplicabilidade no melhoramento genético vegetal. Por fim, o Capítulo V examina as implicações éticas envolvidos na utilização de CRISPR-Cas9 na edição genómica, considerando questões relacionadas com a agricultura, segurança alimentar, impactos ambientais e socioeconómicos, bem como a aceitação pública e as políticas regulatórias em diferentes países. A Conclusão, reafirma a importância da tecnologia CRISPR-Cas9 no avanço do melhoramento vegetal, ao mesmo tempo que destaca a necessidade de mais estudos e debates para garantir o seu uso responsável e ético, considerando os desafios e as oportunidades para a agricultura global.
- CRISPR/Cas9 a simple, inexpensive and effective technique for gene editingPublication . Ferreira, Patrick; Choupina, AltinoIn recent years, the number of tools and techniques that enable genetic material to be added, removed or altered at specifc locations in the genome has increased signifcantly. The objective is to know the structure of genomes, the function of genes and improve gene therapy. In this work we intend to explain the functioning of the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palin dromic Repeats/CRISPR associated protein 9) and the advantages that this technique may have compared to previously developed techniques, such as RNA interference (RNAi), Zinc Finger Nucleases (ZFNs) and transcription activator-like efector nucleases (TALENs) in gene and genome editing. We will start with the story of the discovery, then its biological function in the adaptive immune system of bacteria against bacteriophage attack, and ending with a description of the mechanism of action and its use in gene editing. We will also discuss other Cas enzymes with great potential for use in genome editing as an alternative to Cas9. CRISPR/Cas9 is a simple, inexpensive, and efective technique for gene editing with multiple applications from the development of functional genomics and epigenetics. This technique will, in the near future, have great applications in the development of cell models for use in medical and pharmaceutical processes, in targeted therapy, and improvement of agricultural and environmental species.
- Post-transcriptional gene silencing of glucanase inhibitor protein in Phytophthora cinnamomiPublication . Ferreira, Patrick; Chahed, Abdessalem; Estevinho, Leticia M.; Seixas, Natália; Costa, Rodrigo Arthur Fonseca; Choupina, AltinoInk disease is considered one of the most significant causes contributing to the decline of chestnut orchards. The reduced yield of Castanea sativa Mill can be attributed to two main species: Phytophthora cinnamomi and Phytophthora cambivora, with the first being the main pathogen responsible for ink disease in Portugal. P. cinnamomi is a highly aggressive and widely distributed plant pathogen, capable of infecting nearly 1000 host species. This oomycete causes substantial economic losses and is accountable for the decline of numerous plant species in Europe and worldwide. To date, no effective treatments are available to combat these pathogens. Given chestnut’s economic and ecological significance, particularly in Portugal, it is crucial to investigate the molecular mechanisms underlying the interaction between Phytophthora species and host plants. This can be achieved through the study of the glucanase inhibitor protein (GIP) produced by P. cinnamomi during infection. The technique of RNA interference (RNAi) was employed to suppress the GIP gene of P. cinnamomi. The resulting transformants, carrying the silenced gene, were used to infect C. sativa, allowing for the assessment of the effects of gene silencing on the plant’s phenotype. Additionally, bioinformatics tools predicted the secretion of GIP protein. The obtained results validate RNAi as a potential alternative tool for studying molecular factors and for controlling and managing P. cinnamomi.
- Regulatory non-coding RNAs in crops health and diseasePublication . Ferreira, Patrick; Casquero, Pedro Antonio; Choupina, AltinoFor many years it was thought that the function of RNA was limited to the process of producing proteins. In recent years, scientific discoveries have been proving the multiple roles of different RNAs in different regulatory mechanisms. These RNA’s are collectively called non-coding RNA’s (ncRNA’s). This review presents the latest advances on the different classes of non-coding RNA’s (ncRNA’s) from their function to mechanisms of action. Special emphasis is given to the long non-coding RNAs as new regulatory elements in eukaryote gene expression and in the processes of epigenetic regulation in plants. We believe that increasing studies of regulatory non-coding RNAs in plants will provide a better understanding of the different types of genes related to crop resistance.