Targeting ie-1 gene by RNAi induces baculoviral resistance in lepidopteran cell lines and in transgenic silkworms.
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.
Kanginakudru
S
S
Laboratory of Molecular Genetics, Centre for DNA Fingerprinting and Diagnostics, ECIL Road, Nacharam, Hyderabad, India.
Royer
C
C
Edupalli
S V
SV
Jalabert
A
A
Mauchamp
B
B
Prasad
S V
SV
Chavancy
G
G
Couble
P
P
Nagaraju
J
J
D000818:Animals; D030801:Animals, Genetically Modified / Q000276:immunology / Q000821:virology*; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics / Q000276:immunology / Q000821:virology*; D002460:Cell Line; D018390:Gene Targeting; D005814:Genes, Viral; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D011679:Pupa / Q000235:genetics / Q000821:virology; D034622:RNA Interference; D014170:Transformation, Genetic; D019076:Transgenes; D014760:Viral Fusion Proteins / Q000032:analysis; D010948:Viral Plaque Assay
D014760:Viral Fusion Proteins