A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity
Plants have evolved defense mechanisms to suppress viral transcription and replication by transcriptional and post-transcriptional gene silencing mediated by virus-derived small interfering RNAs (vsiRNAs). Based on this response, virus-induced gene silencing (VIGS)-based technology has been develope...
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MDPI AG
2022-05-01
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| Online Access: | https://www.mdpi.com/2076-3417/12/11/5329 |
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| author | Hernán de Jesús Villanueva-Alonzo Ana Paulina Haro-Álvarez Arturo A. Alvarado-Segura Raúl Enrique Valle-Gough Juan Gualberto Collí-Mull Alberto Cal-Torres Víctor Ermilo Arana-Argáez Julio César Torres-Romero Oscar Alberto Moreno-Valenzuela Geovanny Nic-Can Benjamín Abraham Ayil-Gutiérrez Karla Y. Acosta-Viana |
| author_facet | Hernán de Jesús Villanueva-Alonzo Ana Paulina Haro-Álvarez Arturo A. Alvarado-Segura Raúl Enrique Valle-Gough Juan Gualberto Collí-Mull Alberto Cal-Torres Víctor Ermilo Arana-Argáez Julio César Torres-Romero Oscar Alberto Moreno-Valenzuela Geovanny Nic-Can Benjamín Abraham Ayil-Gutiérrez Karla Y. Acosta-Viana |
| author_sort | Hernán de Jesús Villanueva-Alonzo |
| collection | DOAJ |
| description | Plants have evolved defense mechanisms to suppress viral transcription and replication by transcriptional and post-transcriptional gene silencing mediated by virus-derived small interfering RNAs (vsiRNAs). Based on this response, virus-induced gene silencing (VIGS)-based technology has been developed to silence target genes on either host plants or insect pests. This mechanism could also be used for the silencing of genes of interest in the medical field. We used the VIGS vector pEuMV-YP:Krt18, which was obtained by inserting the <i>Mus musculus</i> (<i>M. musculus</i>) Krt18 sequence into pEuMV-YP:ΔAV1. The objective was to evaluate the capacity of pEuMV-YP:Krt18 to induce <i>Nicotiana benthamiana</i> (<i>N. benthamiana</i>) production of vsiRNAs of a specific sequence that belongs to neither the plant genome nor the wild virus genome, which were used to induce cross-kingdom gene silencing between plants and mammals. The percentage of vsiRNA for each viral gene was calculated from an sRNA library of <i>N. benthamiana</i> plants infected by pEuMV-YP: Krt18. When the vsiRNAs were characterized, it was found that they corresponded to all the genes of the pEuMV-YP:Krt18 vector. These vsiRNAs induced the silencing of the Krt18 gene in <i>M. musculus</i> macrophages, supporting the ability to use VIGS vectors in plants as biofactories for the production of sRNAs that induce gene silencing in mammals. |
| first_indexed | 2024-03-10T01:32:40Z |
| format | Article |
| id | doaj.art-76cb2fadcf744e8d834253e7260354bd |
| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T01:32:40Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-76cb2fadcf744e8d834253e7260354bd2023-11-23T13:39:50ZengMDPI AGApplied Sciences2076-34172022-05-011211532910.3390/app12115329A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing CapacityHernán de Jesús Villanueva-Alonzo0Ana Paulina Haro-Álvarez1Arturo A. Alvarado-Segura2Raúl Enrique Valle-Gough3Juan Gualberto Collí-Mull4Alberto Cal-Torres5Víctor Ermilo Arana-Argáez6Julio César Torres-Romero7Oscar Alberto Moreno-Valenzuela8Geovanny Nic-Can9Benjamín Abraham Ayil-Gutiérrez10Karla Y. Acosta-Viana11Centro de Investigaciones Regionales “Dr. Hideyo Noguchi”, CONACyT-Universidad Autónoma de Yucatán, Av. Itzáes, núm. 490 x calle 59, col. Centro, Mérida 97000, Yucatán, MexicoInstituto de Investigaciones en Ciencias Veterinarias, Universidad Autónoma de Baja California, Carretera, Mexicali-San Felipe Km 3.5, Laguna Campestre, Mexicali 21705, Baja California, MexicoInstituto Tecnológico Superior del Sur del Estado de Yucatán, Tecnológico Nacional de México, Carretera Muna-Felipe Carrillo Puerto Tramo Oxkutzcab-Akil km 41+400, Oxkutzcab 97880, Yucatán, MexicoInstituto de Ciencias Agrícolas, Universidad Autónoma de Baja California, sin núm., Ejido Nuevo León, Mexicali 21705, Baja California, MexicoInstituto Tecnológico Superior de Irapuato, Tecnológico Nacional de México, Carretera Irapuato—Silao km 12.5 Colonia El Copal, Irapuato 36580, Guanajuato, MexicoCentro de Investigaciones Regionales “Dr. Hideyo Noguchi”, Universidad Autónoma de Yucatán, Av. Itzáes, núm. 490 x calle 59, col. Centro, Mérida 97000, Yucatán, MexicoFacultad de Química, Universidad Autónoma de Yucatán, Calle 43 No. 613 x Calle 90, Col. Inalámbrica, Mérida 97069, Yucatán, MexicoFacultad de Química, Universidad Autónoma de Yucatán, Calle 43 No. 613 x Calle 90, Col. Inalámbrica, Mérida 97069, Yucatán, MexicoUnidad de Bioquímica, Centro de Investigación Científica de Yucatán, A.C., Calle 43 No. 130. Col. Chuburná de Hidalgo, Mérida 97000, Yucatán, MexicoFacultad de Ingeniería Química, CONACyT-Universidad Autónoma de Yucatán, Periférico Norte, Km. 33.5. Tablaje Catastral 13615. Col. Chuburná de Hidalgo Inn., Mérida 97203, Yucatán, MexicoCentro de Biotecnología Genómica, CONACyT-Instituto Politécnico Nacional, Blvd. del Maestro, sin núm. esquina, Reynosa 88710, Tamaulipas, MexicoCentro de Investigaciones Regionales “Dr. Hideyo Noguchi”, Universidad Autónoma de Yucatán, Av. Itzáes, núm. 490 x calle 59, col. Centro, Mérida 97000, Yucatán, MexicoPlants have evolved defense mechanisms to suppress viral transcription and replication by transcriptional and post-transcriptional gene silencing mediated by virus-derived small interfering RNAs (vsiRNAs). Based on this response, virus-induced gene silencing (VIGS)-based technology has been developed to silence target genes on either host plants or insect pests. This mechanism could also be used for the silencing of genes of interest in the medical field. We used the VIGS vector pEuMV-YP:Krt18, which was obtained by inserting the <i>Mus musculus</i> (<i>M. musculus</i>) Krt18 sequence into pEuMV-YP:ΔAV1. The objective was to evaluate the capacity of pEuMV-YP:Krt18 to induce <i>Nicotiana benthamiana</i> (<i>N. benthamiana</i>) production of vsiRNAs of a specific sequence that belongs to neither the plant genome nor the wild virus genome, which were used to induce cross-kingdom gene silencing between plants and mammals. The percentage of vsiRNA for each viral gene was calculated from an sRNA library of <i>N. benthamiana</i> plants infected by pEuMV-YP: Krt18. When the vsiRNAs were characterized, it was found that they corresponded to all the genes of the pEuMV-YP:Krt18 vector. These vsiRNAs induced the silencing of the Krt18 gene in <i>M. musculus</i> macrophages, supporting the ability to use VIGS vectors in plants as biofactories for the production of sRNAs that induce gene silencing in mammals.https://www.mdpi.com/2076-3417/12/11/5329vsiRNAcross-kingdomVIGSgene silencing |
| spellingShingle | Hernán de Jesús Villanueva-Alonzo Ana Paulina Haro-Álvarez Arturo A. Alvarado-Segura Raúl Enrique Valle-Gough Juan Gualberto Collí-Mull Alberto Cal-Torres Víctor Ermilo Arana-Argáez Julio César Torres-Romero Oscar Alberto Moreno-Valenzuela Geovanny Nic-Can Benjamín Abraham Ayil-Gutiérrez Karla Y. Acosta-Viana A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity Applied Sciences vsiRNA cross-kingdom VIGS gene silencing |
| title | A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity |
| title_full | A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity |
| title_fullStr | A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity |
| title_full_unstemmed | A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity |
| title_short | A Method to Produce vsiRNAs in Plants with Cross-Kingdom Gene Silencing Capacity |
| title_sort | method to produce vsirnas in plants with cross kingdom gene silencing capacity |
| topic | vsiRNA cross-kingdom VIGS gene silencing |
| url | https://www.mdpi.com/2076-3417/12/11/5329 |
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