Delivery of recombinant adeno-associated virus by jet injection.
The jet-injection technology was used for delivery of recombinant adeno-associated virus (rAAV). Although AAV-based vectors are an attractive tool in gene therapy, some methodological and technical problems of their targeted delivery remain to be solved. We tried to address some of these cell-targeting problems by using a new low-volume needleless injection device the Swiss Injector. First we tested, by electron microscopy, whether jet-injection would have any detrimental effect on rAAV particle integrity. Second, we compared transgene expression after infection of 293T cells with fired or control (non-fired) rAAV that expressed the green fluorescent protein (GFP), beta-galactosidase (beta-gal), the B7.1 molecule, and interleukin 2 (IL2). Third, an rAAV carrying the genes coding for beta-gal was jet-injected into mouse subcutaneous (s.c.) tumours. The staining of tumour cryosections revealed beta-gal expression 72 h after the delivery. Our study demonstrated the applicability of the Swiss Injector for the delivery of rAAV into tumour tissue without either vector particle integrity or the level of expression of the transgenes, as tested in vitro, being affected. The jet-injection technology could improve the distribution of vector particles in the tumour mass without leakage of liquid from the injection site.
Janouskova
Olga
O
Institute of Hematology and Blood Transfusion, U nemocnice 1, Prague 2, 128 20, Czech Republic. olga@uhkt.cz
Nellessen
Thomas
T
Stokrova
Jitka
J
Jinoch
Pavel
P
Smahel
Michal
M
D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006801:Humans; D007277:Injections, Jet; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D004274:DNA, Recombinant; D007376:Interleukin-2; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase