Inhibition of BmNPV replication in silkworm cells using inducible and regulated artificial microRNA precursors targeting the essential viral gene lef-11.
Bombyx mori nucleopolyhedrovirus (BmNPV) is a major silkworm pathogen, causing substantial economic losses to the sericulture industry annually. We demonstrate a novel anti-BmNPV system expressing mature artificial microRNAs (amiRNAs) targeting the viral lef-11 gene. The mature amiRNAs inhibited the lef-11 gene in silkworm BmN-SWU1 cells. Antiviral assays demonstrated that mature amiRNAs silenced the gene and inhibited BmNPV proliferation efficiently. As constitutive overexpression of mature amiRNAs may induce acute cellular toxicity, we further developed a novel virus-induced amiRNA expression system. The amiRNA cassette is regulated by a baculovirus-induced fusion promoter. This baculovirus-induced RNA interference system is strictly regulated by virus infection, which functions in a negative feedback loop to activate the expression of mature amiRNAs against lef-11 and subsequently control inhibition of BmNPV replication. Our study advances the use of a regulatable amiRNA cassette as a safe and effective tool for research of basic insect biology and antiviral application.
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Jun
J
Zhang
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Qian
Q
He
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China; Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016, China.
Chun-Dong
CD
Zhang
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Xiang-Yun
XY
Chen
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Xue-Mei
XM
Chen
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Zhan-Qi
ZQ
Dong
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Na
N
Li
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Xiu-Xiu
XX
Kuang
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
Ming-Ya
MY
Cao
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing 400716, China. Electronic address: lucheng@swu.edu.cn.
Cheng
C
Lu
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing 400716, China. Electronic address: pmh0476@hotmail.com.
Min-Hui
MH
Pan
D000818:Animals; D016367:Baculoviridae / Q000502:physiology*; D002460:Cell Line; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D005814:Genes, Viral; D005822:Genetic Vectors; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
D035683:MicroRNAs; D012322:RNA Precursors
Artificial microRNA; Baculovirus-induced RNA interference system; BmNPV; RNA interference