Delivery of chitosan/dsRNA nanoparticles for silencing of wing development vestigial (vg) gene in Aedes aegypti mosquitoes

被引:82
作者
Kumar, D. Ramesh [1 ,2 ]
Kumar, P. Saravana [2 ]
Gandhi, M. Rajiv [3 ]
Al-Dhabi, Naif Abdullah [4 ]
Paulraj, M. Gabriel [3 ]
Ignacimuthu, S. [3 ,5 ]
机构
[1] Cent Inst Brackishwater Aquaculture, Aquat Anim Hlth & Environm Div, Madras 600028, Tamil Nadu, India
[2] Loyola Coll, Div Microbiol & Mol Biol, Entomol Res Inst, Chennai 600034, Tamil Nadu, India
[3] Loyola Coll, Entomol Res Inst, Madras 600034, Tamil Nadu, India
[4] King Saud Univ, Dept Bot & Microbiol, Coll Sci, Riyadh 11451, Saudi Arabia
[5] King Saud Univ, Coll Sci, Visiting Prof Program, Sci Res, Riyadh 11451, Saudi Arabia
关键词
Chitosan/dsRNA nanoparticles; Vestigial gene (vg); DOUBLE-STRANDED-RNA; IN-VITRO; SIRNA DELIVERY; INTERFERENCE; LARVAL; ULTRABITHORAX; PERSPECTIVES; RESISTANCE; ORIGIN; SYSTEM;
D O I
10.1016/j.ijbiomac.2016.01.030
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) has been used as a gene silencing strategy by the introduction of long double stranded RNA (dsRNA) for the control of pest insects. The aim of the present study was to examine whether the expression of vg gene which is responsible for wing development, can be repressed by chitosan/dsRNA based nanoparticles in Aedes aegypti. The vestigial gene (vg) was amplified from adult mosquito and cloned in pLitmus28i vector. Genetically engineered recombinant plasmid was transformed into RNase III deficient strain for synthesis of bacterially expressed dsRNA. Nanoparticles were prepared via electrostatic interaction between cationic polymer chitosan and anionic nucleic acids (dsRNA). The formation of chitosan/dsRNAnanoparticles and their size were confirmed by Atomic force microscopy (AFM). Chitosan/dsRNA mediated knockdown of Enhanced Green Fluorescence Protein (EGFP) was demonstrated in Sf21 cells. Further, we tested whether such an approach could be used to target vg gene in Ae. aegypti. The results showed that chitosanidsRNA caused significant mortality, delayed growth development and caused adult wing-malformation. A qRT-PCR analysis confirmed that the chitosan/dsRNA mediated transcriptional level was downregulated. Our findings suggest that vg gene intervention strategies through RNAi can emerge as viable option for pest control. (C) 2016 Elsevier B.V. All rights reserved.
引用
收藏
页码:89 / 95
页数:7
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