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2009. Nonviral gene delivery: principle, limitations, and recent progress. Aaps J 11(4):671-81. Alberts AW, Ferguson K, Hennessy S, Vagelos PR. 1974. Regulation of lipid synthesis in cultured animal cells. J Biol Chem 249(16):5241-9. Ansorge S, Lanthier S, Transfiguracion J, Durocher Y, Henry O, Kamen A. 2009. Development of a scalable process for high-yield lentiviral vector production by transient transfection of HEK293 suspension cultures. J Gene Med 11(10):868-76. Balaggan KS, Binley K, Esapa M, Iqball S, Askham Z, Kan O, Tschernutter M, Bainbridge JW, Naylor S, Ali RR.

2000) (Cosset et al. 1995) (Cosset et al. 1995) (Cosset et al. 1995) (Cosset et al. 1995) (Pizzato et al. 2001) (Davis et al. 1997) (Farson et al. 1999) MLV (Swift et al. based 2001) MLV (Schucht et al. based 2006) 3rd with MLV (Coroadinha et RMCE1 based al. 2006b) technology MLV (Loew et al. based 2009) Table 1. ) 20 Viral Gene Therapy Retroviral vectors have been based on several viruses including avian, simian, feline and murine retroviruses, being the latter (MLV) the most used. As so, the majority of the retroviral vector packaging cell lines established were murine derived, being NIH/3T3 the most widely employed.

The envelope cassette encodes typically, but not necessarily, for VSV-G envelope glycoprotein. The development of a fourth generation of lentiviral vectors, rev independent, has also been claimed by means of replacing RRE (rev responsive element) with heterologous viral sequences or by codon-optimization (Bray et al. 1994; Delenda 2004; Kotsopoulou et al. 2000; Pandya et al. 2001; Roberts and Boris-Lawrie 2000). However, its use is not widespread 22 Viral Gene Therapy since, contrary to the other generations of lentiviral vectors, these packaging systems have not been made available for the research community; also the reported titers are typically one to two logs bellow the maximum titers obtained with the second or third generation systems.

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