Sonoporation mediated transduction of pDNA/siRNA into joint synovium in vivo.
The purpose of this study was to investigate the usefulness of sonoporation method on in vivo transduction of plasmid DNA (pDNA) and small interfering RNA (siRNA) into joint tissue. pGEG.GL3 plasmid was mixed with microbubble and injected into knee joints of rats. Ultrasound sonication was performed percutaneously. Three days after injection, GL3 expression of synovial tissue was determined by luciferase assay and RT-PCR. siRNA specific for GL3 (siGL3) or nonspecific siRNA were mixed with pGEG.GL3 plasmid and transduced by sonoporation. siRNA specific for EGFP (siEGFP) was transduced into the knee joints of EGFP transgenic rats, and gene silencing effects for endogenous gene were examined. To determine the localization of transduced siRNA, fluorescently labeled siRNA was transduced into joints. The expression of GL3 in the synovium was significantly enhanced by sonoporation. The gene expression was only seen in the synovium of the knee joint. The expression of GL3 was remarkably suppressed by co-transduction of siGL3, but not suppressed by nonspecific siRNA. siEGFP transduced by sonoporation attenuated green fluorescence on the surface layer of synovium of EGFP transgenic rats. The fluorescently labeled siRNA was seen in the synovium around the patella, femur, and tibia. Sonoporation is examined as a recent, novel, gene transduction method, and the advantage of this technique is minimal invasiveness. In this study, we showed that pDNA/siRNA can be transduced specifically into the joint synovium using sonoporation. The present method may be useful in nucleic acid therapy for joint disorders.
Saito
Masazumi
M
Department of Orthopaedics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
Mazda
Osam
O
Takahashi
Kenji A
KA
Arai
Yuji
Y
Kishida
Tsunao
T
Shin-Ya
Masaharu
M
Inoue
Atsuo
A
Tonomura
Hitoshi
H
Sakao
Kei
K
Morihara
Toru
T
Imanishi
Jiro
J
Kawata
Mitsuhiro
M
Kubo
Toshikazu
T
D000818:Animals; D004247:DNA / Q000235:genetics*; D005786:Gene Expression Regulation; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000378:metabolism; D006614:Hindlimb; D008297:Male; D010957:Plasmids / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013010:Sonication; D013264:Stifle / Q000378:metabolism / Q000473:pathology; D013583:Synovial Membrane / Q000378:metabolism*; D014161:Transduction, Genetic; D019076:Transgenes
D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D004247:DNA