Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects

Jayendra Nath Shukla; Megha Kalsi; Amit Sethi; Kenneth E. Narva; Elane Fishilevich; Satnam Singh; Kanakachari Mogilicherla; Subba Reddy Palli

doi:10.1080/15476286.2016.1191728

Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects

Jayendra Nath Shukla, Megha Kalsi, Amit Sethi, Kenneth E. Narva, Elane Fishilevich, Satnam Singh, Kanakachari Mogilicherla, Subba Reddy Palli

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

196 Scopus citations

Abstract

RNA interference (RNAi) has become a widely used reverse genetic tool to study gene function in eukaryotic organisms and is being developed as a technology for insect pest management. The efficiency of RNAi varies among organisms. Insects from different orders also display differential efficiency of RNAi, ranging from highly efficient (coleopterans) to very low efficient (lepidopterans). We investigated the reasons for varying RNAi efficiency between lepidopteran and coleopteran cell lines and also between the Colorado potato beetle, Leptinotarsa decemlineata and tobacco budworm, Heliothis virescens. The dsRNA either injected or fed was degraded faster in H. virescens than in L. decemlineata. Both lepidopteran and coleopteran cell lines and tissues efficiently took up the dsRNA. Interestingly, the dsRNA administered to coleopteran cell lines and tissues was taken up and processed to siRNA whereas the dsRNA was taken up by lepidopteran cell lines and tissues but no siRNA was detected in the total RNA isolated from these cell lines and tissues. The data included in this paper showed that the degradation and intracellular transport of dsRNA are the major factors responsible for reduced RNAi efficiency in lepidopteran insects.

Original language	English
Pages (from-to)	656-669
Number of pages	14
Journal	RNA Biology
Volume	13
Issue number	7
DOIs	https://doi.org/10.1080/15476286.2016.1191728
State	Published - Jul 2 2016

Bibliographical note

Publisher Copyright:
© 2016 Taylor & Francis Group, LLC.

Funding

This material is based upon work supported by the National Science Foundation I/UCRC, the Center for Arthropod Management Technologies under Grant No. IIP-1338775 and by industry partners. This is publication number 16-08-065 from the Kentucky Agricultural Experimental Station and published with the approval of the director. This work is supported by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000. Special thanks to Dr. Cynthia Goodman for providing Lepd- SL1 and TcA cells and Dr. Seth DeBolt for the use of confocal microscope purchased by KY NSF EPSCoR via National Science Foundation award 1355438.

Funders	Funder number
UCRC
National Science Foundation (NSF)	1355438
U.S. Department of Agriculture	2351177000
National Institute of Food and Agriculture
Kentucky Agricultural Experiment Station
Kansas NSF EPSCoR
Center for Arthropod Management Technologies	IIP-1338775, 16-08-065

Keywords

Coleoptera
RNA interference
double-stranded RNA
lepidoptera

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1080/15476286.2016.1191728

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abstract = "RNA interference (RNAi) has become a widely used reverse genetic tool to study gene function in eukaryotic organisms and is being developed as a technology for insect pest management. The efficiency of RNAi varies among organisms. Insects from different orders also display differential efficiency of RNAi, ranging from highly efficient (coleopterans) to very low efficient (lepidopterans). We investigated the reasons for varying RNAi efficiency between lepidopteran and coleopteran cell lines and also between the Colorado potato beetle, Leptinotarsa decemlineata and tobacco budworm, Heliothis virescens. The dsRNA either injected or fed was degraded faster in H. virescens than in L. decemlineata. Both lepidopteran and coleopteran cell lines and tissues efficiently took up the dsRNA. Interestingly, the dsRNA administered to coleopteran cell lines and tissues was taken up and processed to siRNA whereas the dsRNA was taken up by lepidopteran cell lines and tissues but no siRNA was detected in the total RNA isolated from these cell lines and tissues. The data included in this paper showed that the degradation and intracellular transport of dsRNA are the major factors responsible for reduced RNAi efficiency in lepidopteran insects.",

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Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects

Abstract

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Funding

Keywords

ASJC Scopus subject areas

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