{"author":[{"full_name":"Moorlach, Benjamin Willem","orcid":"0000-0002-7201-4335","orcid_put_code_url":"https://api.orcid.org/v2.0/0000-0002-7201-4335/work/175881812","last_name":"Moorlach","id":"243413","first_name":"Benjamin Willem"},{"first_name":"Désirée","full_name":"Jakobs-Schönwandt, Désirée","last_name":"Jakobs-Schönwandt"},{"first_name":"Minna","last_name":"Poranen","full_name":"Poranen, Minna"},{"first_name":"Maria José","full_name":"Ladera Carmona, Maria José","last_name":"Ladera Carmona"},{"first_name":"Mohamed","last_name":"Abdeldayem","full_name":"Abdeldayem, Mohamed"},{"full_name":"Kogel, Karl-Heinz","last_name":"Kogel","first_name":"Karl-Heinz"},{"first_name":"Ana","last_name":"Sede","full_name":"Sede, Ana"},{"last_name":"Heinlein","full_name":"Heinlein, Manfred","first_name":"Manfred"},{"first_name":"Anant","id":"201870","full_name":"Patel, Anant","last_name":"Patel"}],"keyword":["SIGS","IPEC","chitosan","alginate","dsRNA","RNAi"],"conference":{"end_date":"2024-09-04","location":"Swansea, Vereinigtes Königreich","name":"New IPM Emerging Opportunities ","start_date":"2024-09-02"},"date_updated":"2025-01-17T10:40:43Z","date_created":"2025-01-13T16:10:25Z","user_id":"220548","related_material":{"link":[{"url":"https://www.newipm.com/programmes","relation":"confirmation"}]},"language":[{"iso":"eng"}],"abstract":[{"lang":"eng","text":"Growing concerns over negative effects of pesticides on the environment and on human health are the cause of an increased demand for environmentally sustainable plant protection agents. A promising alternative to conventional chemical pesticides is double-stranded RNA, a nature-derived and environmentally safe molecule that occurs in all organisms and is rapidly degraded in soils. Its application triggers RNA-induced gene silencing, also known as RNA interference (RNAi), an ancient cellular mechanism of eukaryotic cells to protect themselves against pathogens, as well as a means of gene regulation.\r\nIn the project BioProtect, we are developing formulations for spray-induced gene silencing (SIGS) with long dsRNA produced in a bacterial host system (Niehl et al., 2018). We formulated the dsRNA in interpolyelectrolyte complexes (IPECs) composed of the biopolymers chitosan and alginate. Our primary objectives were to enhance stability of dsRNA against nuclease digestion and to mask the negative charge of the dsRNA, which may promote foliar uptake, i.e., overcome plant barriers, such as the cuticle, cell wall and cell membrane. Our approach relies on utilising low polymer concentrations to generate submicron particles, with their charge being either positive or negative determined by the charge ratio during the formulation process. This method incorporates the dsRNA throughout the particle while integrating the nature-derived metal ion chelator, alginate. This serves to inhibit nucleases, which are dependent of metal co-factors. Following this approach, we obtained a high formulation efficiency of > 94 %. Next, we investigated the influence of the charge ratio of positively to negatively charged groups and total polymer content during the formulation process on the size, size distribution and ζ potential of the IPECs. We discovered that formulating dsRNA with alginate and chitosan at low concentrations ≤ 0.05 g/L produced < 100 nm particles. Moreover, we obtained submicron IPECs at charge ratios ≤ 0.9 (+/-) and ≥ 1.25 (+/-), exhibiting negative and positive ζ potentials, respectively. Furthermore, the IPEC formulation protected the dsRNA from enzymatic degradation by nucleases. In addition, we observed outstanding protection of formulated dsRNA from heat degradation. Experiments on Nicotiana benthamiana plants showed that heat-treated dsRNA formulations offered protection against Tobacco mosaic virus compared to unformulated dsRNA. Further experiments with shorter (448 bp) dsRNA targeting the rice blast fungus Magnaporthe oryzae demonstrated that our dsRNA-formulations significantly reduced the necrotic area and fungal mass in Brachypodium distachyon. In essence, this formulation demonstrates versatility and adaptability for the production of IPECs with customizable size, surface charge, and nucleic acid content. The incorporation of target-specific dsRNA sequences into this formulation pipeline holds promise for streamlining the large-scale implementation of non-transformative RNAi-based crop protection, such as spray-induced gene silencing.\r\n"}],"_id":"5310","status":"public","citation":{"apa":"Moorlach, B. W., Jakobs-Schönwandt, D., Poranen, M., Ladera Carmona, M. J., Abdeldayem, M., Kogel, K.-H., … Patel, A. (2024). Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection. Presented at the New IPM Emerging Opportunities , Swansea, Vereinigtes Königreich.","ama":"Moorlach BW, Jakobs-Schönwandt D, Poranen M, et al. Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection. In: ; 2024.","ieee":"B. W. Moorlach <i>et al.</i>, “Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection,” presented at the New IPM Emerging Opportunities , Swansea, Vereinigtes Königreich, 2024.","bibtex":"@inproceedings{Moorlach_Jakobs-Schönwandt_Poranen_Ladera Carmona_Abdeldayem_Kogel_Sede_Heinlein_Patel_2024, title={Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection}, author={Moorlach, Benjamin Willem and Jakobs-Schönwandt, Désirée and Poranen, Minna and Ladera Carmona, Maria José and Abdeldayem, Mohamed and Kogel, Karl-Heinz and Sede, Ana and Heinlein, Manfred and Patel, Anant}, year={2024} }","short":"B.W. Moorlach, D. Jakobs-Schönwandt, M. Poranen, M.J. Ladera Carmona, M. Abdeldayem, K.-H. Kogel, A. Sede, M. Heinlein, A. Patel, in: 2024.","mla":"Moorlach, Benjamin Willem, et al. <i>Formulation of in Vivo Produced Long Double-Stranded RNA for Plant Protection</i>. 2024.","chicago":"Moorlach, Benjamin Willem, Désirée Jakobs-Schönwandt, Minna Poranen, Maria José Ladera Carmona, Mohamed Abdeldayem, Karl-Heinz Kogel, Ana Sede, Manfred Heinlein, and Anant Patel. “Formulation of in Vivo Produced Long Double-Stranded RNA for Plant Protection,” 2024.","alphadin":"<span style=\"font-variant:small-caps;\"><span style=\"font-variant:small-caps;\">Moorlach, Benjamin Willem</span> ; <span style=\"font-variant:small-caps;\">Jakobs-Schönwandt, Désirée</span> ; <span style=\"font-variant:small-caps;\">Poranen, Minna</span> ; <span style=\"font-variant:small-caps;\">Ladera Carmona, Maria José</span> ; <span style=\"font-variant:small-caps;\">Abdeldayem, Mohamed</span> ; <span style=\"font-variant:small-caps;\">Kogel, Karl-Heinz</span> ; <span style=\"font-variant:small-caps;\">Sede, Ana</span> ; <span style=\"font-variant:small-caps;\">Heinlein, Manfred</span> ; u. a.</span>: Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection. In: , 2024"},"title":"Formulation of in vivo Produced Long Double-stranded RNA for Plant Protection","type":"conference","year":"2024"}