Grave L, Dreyer D, Dieterle A, Leroy P, Michou A I, Doderer C, Pavirani A, Lusky M, Mehtali M
Transgène SA, Strasbourg, France.
J Gene Med. 2000 Nov-Dec;2(6):433-43. doi: 10.1002/1521-2254(200011/12)2:6<433::AID-JGM143>3.0.CO;2-1.
Strong and stable transgene expression is fundamental to the success of recombinant adenovirus vectors in human gene therapy. However, control of transgene expression is a complex process, involving both viral and cellular factors. In this study, the influence of the E4 adenoviral region on the activity of various promoters was investigated in vitro and in vivo.
Pairs of isogenic E1o and E1oE4o vectors were generated and compared. Levels of transgene expression were determined by Northern blot, ELISA and FACS analysis. Initiation of transcription was studied by nuclear run-on assays.
Similar to the viral CMV and RSV promoters, the activity of the ubiquitous cellular PGK promoter required the presence of the E4 genes in vitro and in vivo. In contrast, transgene expression from selected liver- and tumor-specific promoters did not require E4 functions.
Together with the reported low liver toxicity of E1oE4o vectors, the independence of E4 of liver-specific promoters renders such vectors interesting alternatives to the use of gutless vectors.
强大而稳定的转基因表达是重组腺病毒载体在人类基因治疗中取得成功的基础。然而，转基因表达的控制是一个复杂的过程，涉及病毒和细胞因素。在本研究中，在体外和体内研究了E4腺病毒区域对各种启动子活性的影响。
构建并比较了成对的同基因E1o和E1oE4o载体。通过Northern印迹、ELISA和FACS分析确定转基因表达水平。通过核转录分析研究转录起始。
与病毒CMV和RSV启动子类似，普遍存在的细胞PGK启动子的活性在体外和体内都需要E4基因的存在。相反，从选定的肝脏和肿瘤特异性启动子的转基因表达不需要E4功能。
连同报道的E1oE4o载体低肝脏毒性，肝脏特异性启动子对E4的独立性使得此类载体成为使用无肠载体的有趣替代方案。
