Plasmid engineering for controlled and sustained gene expression for nonviral gene therapy.
Gene therapy requires the introduction of genetic material in diseased cells with the aim of treating or ultimately curing a disease. Since the start of gene therapy clinical trials in 1990, gene therapy has proven to be possible, but studies to date have highlighted the difficulty of achieving efficient, specific, and long-term transgene expression. Efforts to improve gene therapy strategies over the past years were mainly aimed at solving the problem of delivery, without paying much attention to the optimization of the expression cassette. With the current understanding of the eukaryotic transcription machinery and advanced molecular biology techniques at our disposition, it has now become possible to create custom-made transgene expression cassettes optimized for gene therapy applications. In this review, we will discuss several strategies that have been explored to improve the level and duration of transgene expression, to increase control over expression, or to restrict transgene expression to specific cell types or tissues. Although still in its infancy, such strategies will eventually lead to improvement of nonviral gene therapy and expansion of the range of possible therapeutic applications.
van Gaal
Ethlinn V B
EV
Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, P.O Box 80082, 3508 TB, Utrecht, The Netherlands.
Hennink
Wim E
WE
Crommelin
Daan J A
DJ
Mastrobattista
Enrico
E
D021581:Active Transport, Cell Nucleus; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis / Q000378:metabolism*; D025461:Feedback, Physiological; D005786:Gene Expression Regulation / Q000187:drug effects; D018390:Gene Targeting; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000639:trends*; D005822:Genetic Vectors / Q000096:biosynthesis*; D006801:Humans; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D019076:Transgenes
D004247:DNA; D013752:Tetracycline