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

被引:21
作者
Chen, Xien [1 ]
Koo, Jinmo [1 ]
Gurusamy, Dhandapani [1 ,2 ]
Mogilicherla, Kanakachari [1 ,3 ]
Reddy Palli, Subba [2 ]
机构
[1] Univ Kentucky, Coll Agr Food & Environm, Dept Entomol, Lexington, KY 40546 USA
[2] Bharathiar Univ, Kongunadu Arts & Sci Coll Autonomous, Dept Bot, Coimbatore, Tamil Nadu, India
[3] ICAR Directorate Poultry Res, Div Mol Genet, Hyderabad, India
基金
美国国家科学基金会;
关键词
RNA interference; dsRNA transporter; enhanced RNAi; transgenesis; Spodoptera frugiperda; GENETIC-TRANSFORMATION; DSRNA; PIGGYBAC; EXPRESSION; TRANSPORT; CELLS; MOTH; L;
D O I
10.1080/15476286.2020.1842632
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:1291 / 1299
页数:9
相关论文
共 35 条
[1]   Beyond Drosophila: RNAi In Vivo and Functional Genomics in Insects [J].
Belles, Xavier .
ANNUAL REVIEW OF ENTOMOLOGY, 2010, 55 :111-128
[2]   The involvement of clathrin-mediated endocytosis and two Sid-1-like transmembrane proteins in doublestranded RNA uptake in the Colorado potato beetle midgut [J].
Cappelle, K. ;
de Oliveira, C. F. R. ;
Van Eynde, B. ;
Christiaens, O. ;
Smagghe, G. .
INSECT MOLECULAR BIOLOGY, 2016, 25 (03) :315-323
[3]   Identification and characterization of highly active promoters from the fall armyworm, Spodoptera frugiperda [J].
Chen, Xien ;
Chereddy, Shankar C. R. R. ;
Gurusamy, Dhandapani ;
Palli, Subba Reddy .
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 2020, 126
[4]   Identification and functional analysis of promoters of heat-shock genes from the fall armyworm, Spodoptera frugiperda [J].
Chen, Xien ;
Tan, Anjiang ;
Palli, Subba Reddy .
SCIENTIFIC REPORTS, 2020, 10 (01)
[5]   Hyperactive piggyBac transposase improves transformation efficiency in diverse insect species [J].
Eckermann, Kolja N. ;
Ahmed, Hassan M. M. ;
KaramiNejadRanjbar, Mohammad ;
Dippel, Stefan ;
Ogaugwu, Christian E. ;
Kitzmann, Peter ;
Isah, Musa D. ;
Wimmer, Ernst A. .
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 2018, 98 :16-24
[6]   Transport of dsRNA into cells by the transmembrane protein SID-1 [J].
Feinberg, EH ;
Hunter, CP .
SCIENCE, 2003, 301 (5639) :1545-1547
[7]   Genetic transformation of the codling moth, Cydia pomonella L., with piggyBac EGFP [J].
Ferguson, Holly J. ;
Neven, Lisa G. ;
Thibault, Stephen T. ;
Mohammed, Ahmed ;
Fraser, Malcolm .
TRANSGENIC RESEARCH, 2011, 20 (01) :201-214
[8]   Small silencing RNAs: an expanding universe [J].
Ghildiyal, Megha ;
Zamore, Phillip D. .
NATURE REVIEWS GENETICS, 2009, 10 (02) :94-108
[9]   A nuclease specific to lepidopteran insects suppresses RNAi [J].
Guan, Ruo-Bing ;
Li, Hai-Chao ;
Fan, Yu-Jie ;
Hu, Shao-Ru ;
Christiaens, Olivier ;
Smagghe, Guy ;
Miao, Xue-Xia .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2018, 293 (16) :6011-6021
[10]   Lipids help double-stranded RNA in endosomal escape and improve RNA interference in the fall armyworm, Spodoptera frugiperda [J].
Gurusamy, Dhandapani ;
Mogilicherla, Kanakachari ;
Shukla, Jayendra Nath ;
Palli, Subba Reddy .
ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 2020, 104 (04)