Kanginakudru S, Royer C, Edupalli S V, Jalabert A, Mauchamp B, Prasad S V, Chavancy G, Couble P, Nagaraju J
Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad, India.
Insect Mol Biol. 2007 Oct;16(5):635-44. doi: 10.1111/j.1365-2583.2007.00753.x.
RNA interference (RNAi)-mediated viral inhibition has been used in a few organisms for eliciting viral resistance. In the present study, we report the use of RNAi in preventing baculovirus infection in a lepidopteran. We targeted the baculoviral immediate early-1 (ie-1) gene in both a transformed lepidopteran cell line and in the transgenic silkworm Bombyx mori L. Constitutive expression of double-stranded RNA was achieved by piggyBac-mediated transformation of Sf9 cell line with a transgene encoding double-stranded ie-1 RNA (dsie-1). Strong viral repression was seen at early stages of infection but subsequent recovery of viral proliferation was observed. In contrast, the same transgene inserted into the chromosomes of transgenic silkworms induced long-term inhibition of B. mori nucleopolyhedrovirus infection, with nearly 40% protection compared with nontransgenic animals. Protection was efficient at larval stages after oral infection with occlusion bodies or hemocoel injection of budded viruses. Virus injected pupae also displayed resistance. These results show that heritable RNAi can be used to protect silkworm strains from baculovirus infection.
RNA干扰(RNAi)介导的病毒抑制已在一些生物体中用于引发病毒抗性。在本研究中,我们报道了利用RNAi预防鳞翅目昆虫中的杆状病毒感染。我们在转化的鳞翅目昆虫细胞系和转基因家蚕(Bombyx mori L.)中靶向杆状病毒立即早期-1(ie-1)基因。通过piggyBac介导的Sf9细胞系转化,用编码双链ie-1 RNA(dsie-1)的转基因实现了双链RNA的组成型表达。在感染早期观察到强烈的病毒抑制,但随后观察到病毒增殖的恢复。相比之下,插入转基因家蚕染色体中的相同转基因诱导了对家蚕核型多角体病毒感染的长期抑制,与非转基因动物相比,保护率近40%。在用包涵体口服感染或经血腔注射芽生病毒后,幼虫阶段的保护是有效的。注射病毒的蛹也表现出抗性。这些结果表明,可遗传的RNAi可用于保护家蚕品系免受杆状病毒感染。