RRC ID 3504
Author Kawashima T, Osanai M, Futahashi R, Kojima T, Fujiwara H.
Title A novel target-specific gene delivery system combining baculovirus and sequence-specific long interspersed nuclear elements.
Journal Virus Res.
Abstract Transposable elements are valuable for somatic and germ-line transformation. However, long interspersed nuclear elements (LINEs) have not been used because of poor information on the transposition mechanism. We have developed a novel gene delivery system combining baculovirus AcNPV and two silkworm LINEs, SART1 and R1, which integrate into specific sequences of telomeric repeats and 28S ribosomal DNA, respectively. When two LINEs containing the enhanced green fluorescent protein gene recombined into AcNPV were infected into fifth instar larvae of the silkworm, we observed target-specific retrotransposition of LINEs at 72h post-infection, using polymerase chain reaction amplification and sequencing. Telomere- and 28S rDNA-specific transposition occurred in all nine tissues tested, including the ovary and testis. This is the first demonstration of site-specific gene delivery in living larvae. Insertion efficiencies were dependent on the virus titer for injection and the host strains of Bombyx mori. Using this system, we successfully detected the intergeneration transmission of retrotransposed sequences. In addition, AcNPV-mediated SART1 also transposed into telomere of another lepidopteran, Orgyia recens, suggesting that this system is useful for a wide variety of AcNPV-infectious insects. Site-specific gene delivery by virus-mediated LINE will be a potential gene therapy tool to avoid harmful unexpected insertions.
Volume 127(1)
Pages 49-60
Published 2007-7
DOI 10.1016/j.virusres.2007.03.014
PII S0168-1702(07)00103-7
PMID 17498830
MeSH Animals Baculoviridae / genetics* Baculoviridae / metabolism Gene Transfer Techniques* Genes, Insect Genetic Vectors Lepidoptera / classification Lepidoptera / genetics* Long Interspersed Nucleotide Elements / genetics* Telomere* / genetics Transcription, Genetic / genetics Transposases
IF 2.736
Times Cited 4
Silkworms p strain ?