Bushman F D
Infectious Disease Laboratory, Salk Institute, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
Curr Top Microbiol Immunol. 2002;261:165-77. doi: 10.1007/978-3-642-56114-6_8.
Retroviruses integrate into naked DNA in a generally sequence nonspecific fashion, but closer study reveals a variety of forces that influence target site selection. Primary sequence of the target plays a small but detectable role. Proteins bound to target DNA can inhibit integration by blocking access of integration complexes or stimulate integration by distorting DNA. An important example of the latter is DNA distortion in nucleosomal DNA. In vivo integration has not yet been convincingly shown to be biased in favor of any identifiable sequence features, though this could still change in future studies. Many applications of retroviral vectors could be facilitated by targeting integration in vivo to predetermined sites. Towards this end, several groups have studied the properties of fusions of integrase proteins to sequence-specific DNA-binding domains. To date such studies establish that targeting can work well in reactions in vitro, but a variety of obstacles complicate applications in vivo. However, naturally occurring retrotransposons do carry out highly targeted integration using retrovirus-like integrase proteins, fueling long-term hopes for targeting with retroviral integrases as well.
逆转录病毒通常以序列非特异性的方式整合到裸露的DNA中,但进一步研究发现有多种力量影响靶位点的选择。靶标的一级序列起的作用虽小但可检测到。与靶DNA结合的蛋白质可通过阻止整合复合物的进入来抑制整合,或通过使DNA变形来刺激整合。后者的一个重要例子是核小体DNA中的DNA变形。体内整合尚未被令人信服地证明偏向于任何可识别的序列特征,不过在未来研究中这仍可能改变。将逆转录病毒载体在体内的整合靶向预定位点可促进其许多应用。为此,几个研究小组研究了整合酶蛋白与序列特异性DNA结合结构域融合体的特性。迄今为止,此类研究表明靶向在体外反应中能很好地发挥作用,但多种障碍使体内应用变得复杂。然而,天然存在的逆转座子确实利用逆转录病毒样整合酶蛋白进行高度靶向的整合,这也为使用逆转录病毒整合酶进行靶向带来了长期希望。