In vivo cell type-specific gene delivery with retroviral vectors that display single chain antibodies

Abstract

Cell type-specific gene delivery will be essential for in vivo display scAs against the human her2neu cell surface progene therapy. Our laboratory has previously developed tein, was injected. Cells were recovered from the peritoretroviral vector particles, derived from spleen necrosis neum, subjected to antibiotic selection, and tested for the virus, SNV, which display the antigen-binding site of an expression of a lacZ gene transduced by the retroviral vecantibody on the viral surface. Such particles infected only tor. We found that human target cells, which express her2-human cells in vitro, which expressed a receptor recogneu, were infected in vivo. However, neither human cells nized by the antibody. To test cell type-specific gene delivthat do not express her2neu, nor normal mouse cells were ery in vivo, a mouse model system has been developed.infected by such viral particles. These data give proof of Antibiotic resistant human target and non-target cells were principle that retroviral vector-mediated, cell type-specific injected into the peritoneum of SCID mice. Subsequently, gene delivery can be obtained in vivo. a vector solution containing 10 6 infectious particles, which

Changing Viral Tropism Using Immunoliposomes Alters the Stability of Gene Expression: Implications for Viral Vector Design

“…Previous groups have also shown short-term expression after transduction with either lentivirus or MMLV. This could be due to transfer of protein by the viral particles (59,60) or, in the case of viruses that have been disabled in such as a way as to prevent either reverse transcription of RNA or integration of virally derived DNA into the host genome, as a result of direct translation from virally delivered RNA (61) or transcription of episomal viral DNA (60,62). Such "pseudotransduction" has been suggested both as a way of transient expression of toxic proteins (61) and as a safe, nonintegrating, viral vector for gene therapy (62).…”

“…For example, fusing a gene for a single chain antibody (sFv) to the region of the envelope gene encoding its N-terminal amino acids has been carried out (63). This technique has been attempted using different sFvs and other proteins capable of specifically attaching to cellular receptors (62). Most have resulted in very low infectivity, less than 3%, and poor virion stability (63).…”

Recombinant single-chain antibodies with various oligopeptide tails for targeted gene delivery

“…1,2 Alternatively, certain biological activities such as ligands, receptors, antigens and antibodies have been challenged to select cell types. [3][4][5][6][7] These novel approaches have made substantial improvement, but yet further innovation has been in demand.…”

Surface-engineering of lentiviral vectors

“…Many of these chimeric glycoproteins fold correctly, are stably incorporated on virions and allow efficient retargeted virion binding to the expected cell-surface molecules. Large, foreign polypeptides have also been fused directly to all or part of the TM [93][94][95][96][97][98]. However, for most N-terminally substituted chimeric envelope glycoproteins that have been studied to date, the efficiency of viral incorporation has been low, or undetected, possibly as a consequence of suboptimal folding, assembly or transport to the cell surface.…”