Caenorhabditis elegans systemic RNA interference defective protein 1 enhances RNAi efficiency in a lepidopteran insect, the fall armyworm, in a tissue-specific manner

Xien Chen; Jinmo Koo; Dhandapani Gurusamy; Kanakachari Mogilicherla; Subba Reddy Palli

doi:10.1080/15476286.2020.1842632

Caenorhabditis elegans systemic RNA interference defective protein 1 enhances RNAi efficiency in a lepidopteran insect, the fall armyworm, in a tissue-specific manner

Xien Chen, Jinmo Koo, Dhandapani Gurusamy, Kanakachari Mogilicherla, Subba Reddy Palli

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

RNA interference (RNAi) is an important tool for gene function studies in insects, especially in non-model insects. This technology is also being developed for pest control. However, variable RNAi efficiency among insects is limiting its use in insects. Systemic RNAi in Caenorhabditis elegans requires systemic RNA interference defective protein 1 (CeSid1). The expression of CeSid1 in insect cell lines was shown to improve RNAi. However, the mechanisms through which this double-stranded RNA (dsRNA) transporter improves RNAi efficiency in insects is not known. We stably expressed CeSid1 in two Spodoptera frugiperda cell lines, Sf9 and Sf17 cells derived from ovary and midgut, respectively. Expression of CeSid1 enhanced RNAi efficiency in ovarian Sf9 cells, but not in midgut Sf17 cells. Reduced accumulation of dsRNA in late endosomes and successful processing dsRNA to siRNA contribute to enhanced RNAi efficiency in Sf9 cells. Transgenic S. frugiperda expressing CeSid1 were produced and tested for RNAi efficiency. RNAi efficiency enhancement due to CeSid1 expression showed tissue specificity. Compared to RNAi efficiency in wild-type S. frugiperda, CeSid1 expressing transgenic S. frugiperda showed a significant improvement of RNAi in tissues such as Verson’s glands. In contrast, no improvement in RNAi was observed in tissues such as midgut. The in vitro cell-type specific and in vivo tissue-specific enhancement of RNAi efficiency by CeSid1 in S. frugiperda provides valuable information for improving RNAi in insects such as those belonging to order Lepidoptera where RNAi is variable and inefficient.

Original language	English
Pages (from-to)	1291-1299
Number of pages	9
Journal	RNA Biology
Volume	18
Issue number	9
DOIs	https://doi.org/10.1080/15476286.2020.1842632
State	Published - 2021

Bibliographical note

Funding Information:
We thank Dr Anjiang Tan of Shanghai institute of plant physiology and ecology, China for the gift of piggyBac and pTD1 vectors, Dr Cindy Goodman from USDA ARS for the gift of Sf17 cells and Jeff Howell from University of Kentucky for help with Fall armyworm rearing The research reported is supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under Grant No IIP‐1821936 and by industry partners, Agriculture and Food Research Initiative Competitive Grant No. 2019-67013-29351 and the National Institute of Food and Agriculture, US Department of Agriculture (2353057000).

Publisher Copyright:
© 2020 Informa UK Limited, trading as Taylor & Francis Group.

Keywords

RNA interference
Spodoptera frugiperda
dsRNA transporter
enhanced RNAi
transgenesis

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1080/15476286.2020.1842632

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title = "Caenorhabditis elegans systemic RNA interference defective protein 1 enhances RNAi efficiency in a lepidopteran insect, the fall armyworm, in a tissue-specific manner",

abstract = "RNA interference (RNAi) is an important tool for gene function studies in insects, especially in non-model insects. This technology is also being developed for pest control. However, variable RNAi efficiency among insects is limiting its use in insects. Systemic RNAi in Caenorhabditis elegans requires systemic RNA interference defective protein 1 (CeSid1). The expression of CeSid1 in insect cell lines was shown to improve RNAi. However, the mechanisms through which this double-stranded RNA (dsRNA) transporter improves RNAi efficiency in insects is not known. We stably expressed CeSid1 in two Spodoptera frugiperda cell lines, Sf9 and Sf17 cells derived from ovary and midgut, respectively. Expression of CeSid1 enhanced RNAi efficiency in ovarian Sf9 cells, but not in midgut Sf17 cells. Reduced accumulation of dsRNA in late endosomes and successful processing dsRNA to siRNA contribute to enhanced RNAi efficiency in Sf9 cells. Transgenic S. frugiperda expressing CeSid1 were produced and tested for RNAi efficiency. RNAi efficiency enhancement due to CeSid1 expression showed tissue specificity. Compared to RNAi efficiency in wild-type S. frugiperda, CeSid1 expressing transgenic S. frugiperda showed a significant improvement of RNAi in tissues such as Verson{\textquoteright}s glands. In contrast, no improvement in RNAi was observed in tissues such as midgut. The in vitro cell-type specific and in vivo tissue-specific enhancement of RNAi efficiency by CeSid1 in S. frugiperda provides valuable information for improving RNAi in insects such as those belonging to order Lepidoptera where RNAi is variable and inefficient.",

keywords = "RNA interference, Spodoptera frugiperda, dsRNA transporter, enhanced RNAi, transgenesis",

author = "Xien Chen and Jinmo Koo and Dhandapani Gurusamy and Kanakachari Mogilicherla and {Reddy Palli}, Subba",

note = "Funding Information: We thank Dr Anjiang Tan of Shanghai institute of plant physiology and ecology, China for the gift of piggyBac and pTD1 vectors, Dr Cindy Goodman from USDA ARS for the gift of Sf17 cells and Jeff Howell from University of Kentucky for help with Fall armyworm rearing The research reported is supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies, under Grant No IIP‐1821936 and by industry partners, Agriculture and Food Research Initiative Competitive Grant No. 2019-67013-29351 and the National Institute of Food and Agriculture, US Department of Agriculture (2353057000). Publisher Copyright: {\textcopyright} 2020 Informa UK Limited, trading as Taylor & Francis Group.",

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AU - Mogilicherla, Kanakachari

AU - Reddy Palli, Subba

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PY - 2021

Y1 - 2021

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Caenorhabditis elegans systemic RNA interference defective protein 1 enhances RNAi efficiency in a lepidopteran insect, the fall armyworm, in a tissue-specific manner

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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