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Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests

Genetically modified (GM) crops producing double-stranded RNAs (dsRNAs) are being investigated largely as an RNA interference (RNAi)-based resistance strategy against crop insect pests. However, limitations of this strategy include the sensitivity of dsRNA to insect gut nucleases and its poor insect...

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Autores principales: Gillet, François-Xavier, Garcia, Rayssa A., Macedo, Leonardo L. P., Albuquerque, Erika V. S., Silva, Maria C. M., Grossi-de-Sa, Maria F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408074/
https://www.ncbi.nlm.nih.gov/pubmed/28503153
http://dx.doi.org/10.3389/fphys.2017.00256
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author Gillet, François-Xavier
Garcia, Rayssa A.
Macedo, Leonardo L. P.
Albuquerque, Erika V. S.
Silva, Maria C. M.
Grossi-de-Sa, Maria F.
author_facet Gillet, François-Xavier
Garcia, Rayssa A.
Macedo, Leonardo L. P.
Albuquerque, Erika V. S.
Silva, Maria C. M.
Grossi-de-Sa, Maria F.
author_sort Gillet, François-Xavier
collection PubMed
description Genetically modified (GM) crops producing double-stranded RNAs (dsRNAs) are being investigated largely as an RNA interference (RNAi)-based resistance strategy against crop insect pests. However, limitations of this strategy include the sensitivity of dsRNA to insect gut nucleases and its poor insect cell membrane penetration. Working with the insect pest cotton boll weevil (Anthonomus grandis), we showed that the chimeric protein PTD-DRBD (peptide transduction domain—dsRNA binding domain) combined with dsRNA forms a ribonucleoprotein particle (RNP) that improves the effectiveness of the RNAi mechanism in the insect. The RNP slows down nuclease activity, probably by masking the dsRNA. Furthermore, PTD-mediated internalization in insect gut cells is achieved within minutes after plasma membrane contact, limiting the exposure time of the RNPs to gut nucleases. Therefore, the RNP provides an approximately 2-fold increase in the efficiency of insect gene silencing upon oral delivery when compared to naked dsRNA. Taken together, these data demonstrate the role of engineered RNPs in improving dsRNA stability and cellular entry, representing a path toward the design of enhanced RNAi strategies in GM plants against crop insect pests.
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spelling pubmed-54080742017-05-12 Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests Gillet, François-Xavier Garcia, Rayssa A. Macedo, Leonardo L. P. Albuquerque, Erika V. S. Silva, Maria C. M. Grossi-de-Sa, Maria F. Front Physiol Physiology Genetically modified (GM) crops producing double-stranded RNAs (dsRNAs) are being investigated largely as an RNA interference (RNAi)-based resistance strategy against crop insect pests. However, limitations of this strategy include the sensitivity of dsRNA to insect gut nucleases and its poor insect cell membrane penetration. Working with the insect pest cotton boll weevil (Anthonomus grandis), we showed that the chimeric protein PTD-DRBD (peptide transduction domain—dsRNA binding domain) combined with dsRNA forms a ribonucleoprotein particle (RNP) that improves the effectiveness of the RNAi mechanism in the insect. The RNP slows down nuclease activity, probably by masking the dsRNA. Furthermore, PTD-mediated internalization in insect gut cells is achieved within minutes after plasma membrane contact, limiting the exposure time of the RNPs to gut nucleases. Therefore, the RNP provides an approximately 2-fold increase in the efficiency of insect gene silencing upon oral delivery when compared to naked dsRNA. Taken together, these data demonstrate the role of engineered RNPs in improving dsRNA stability and cellular entry, representing a path toward the design of enhanced RNAi strategies in GM plants against crop insect pests. Frontiers Media S.A. 2017-04-28 /pmc/articles/PMC5408074/ /pubmed/28503153 http://dx.doi.org/10.3389/fphys.2017.00256 Text en Copyright © 2017 Gillet, Garcia, Macedo, Albuquerque, Silva and Grossi-de-Sa. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Gillet, François-Xavier
Garcia, Rayssa A.
Macedo, Leonardo L. P.
Albuquerque, Erika V. S.
Silva, Maria C. M.
Grossi-de-Sa, Maria F.
Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title_full Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title_fullStr Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title_full_unstemmed Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title_short Investigating Engineered Ribonucleoprotein Particles to Improve Oral RNAi Delivery in Crop Insect Pests
title_sort investigating engineered ribonucleoprotein particles to improve oral rnai delivery in crop insect pests
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408074/
https://www.ncbi.nlm.nih.gov/pubmed/28503153
http://dx.doi.org/10.3389/fphys.2017.00256
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