Enhanced in vitro and in vivo gene delivery using cationic agent complexed retrovirus vectors

Abstract

Retroviruses are, at present, the most efficient integrative mediated infection without cytotoxicity. To increase virus vectors available for gene delivery. However, these viruses infectivity further we combined the enhancing effect of are still limited by relatively low titres. Although several pro-DOGS on virus infectivity with concentration of virus partocols exist to improve virus titre most of them are timeticles by ultrafiltration to reach titres of 1 × 10 9 IU/ml. The consuming and unable to provide sufficient virus for in vivo in vivo transduction of regenerating rat liver, by an amphoapplications. Virus titre can be enhanced by polybrene and tropic retrovirus was increased approximately five-fold by other cationic agents. By investigating a broad range of the addition of DOGS compared with virus alone. There cationic agents for their ability to enhance virus infectivity was no animal toxicity observed following the adminiswe found that both ecotropic and amphotropic retrovirus tration of DOGS. The improved transduction efficiency infection could be increased. In particular, the lipopolyseen both in vitro and in vivo following the co-adminisamine dioctadecylamidoglycylspermine (DOGS) gave up to tration of DOGS/virus complexes may be useful for future one order of magnitude enhancement above polybrenegene therapy applications.

Efficient Enhancement of Lentiviral Transduction Efficiency in Murine Spermatogonial Stem Cells

“…Indeed, when polybrene 4 ug/ml and DOGS 2 ug/ml were microinjected into rat testes, and the testes were histologically analyzed after 2-5 months, DOGS-treated group displayed spermatogenesis in the seminiferous tubules of more than 80% of the recipients while in the polybrene-treated groups showed spermatogenesis in only 20% of the recipients (unpublished data). Themis and Forbes (1998) performed internal transduction by injecting retrovirus directly into the body, and showed efficient transduction into tissue cells can be achieved by the addition of DOGS. Similarly our results for this study showed lower cytotoxicity for the DOGS-treated group compared with the polybrene-treated group (Fig.…”

Tri-iodothyronine and a deleted form of hepatocyte growth factor act synergistically to enhance liver proliferation and enable in vivo retroviral gene transfer via the peripheral venous system

“…During the resultant regeneration phase the liver can be efficiently transduced with retroviral vectors, achieving hepatocyte transduction efficiencies of up to 35% following a single injection of vector. [9][10][11][12] Unfortunately, PH is a clinically unacceptable technique. Another approach has been to occlude a branch of the portal vein.…”

“…This enabled a four-fold increase in the in vivo transduction efficiency of the post-hepatectomy liver. 12 In this study we combined the administration of two growth factors with disparate intracellular mechanisms to induce hepatocyte proliferation in the intact liver and enable transduction with the amphotropic virus vector TELCeB/AF-7 21 complexed with the cationic liposome DOGS.…”

A novel human suspension culture packaging cell line for production of high–titre retroviral vectors

“…Virus containing supernatants were titred after filtration (0.8 m filters; Sartorius, Epsom, UK) in the presence of the polycation polybrene at 4 g/ml or the polyamine DOGS (dioctadecyl dimethyl ammonium chloride -Transfectam; Promega, Southampton, UK) at 5 g/ml as previously described. 21 Vector production under reduced serum conditions Virus producing WIL-E cells were grown at 6 × 10 6 cells/ml in RPMI 1640 medium or complete suspension medium (CSM -RPMI 1640 medium supplemented with 15% serum and 20% HL-1 medium) then transferred to medium with or without serum ( Figure 5). Supernatants were harvested after 18 h then titred on NIH3T3 cells.…”