慢病毒载体介导的转基因整合位点研究方法

慢病毒载体整合到宿主细胞的染色体上可以长期稳定表达,目前已成为基因治疗和转基因动物载体研究的热点.慢病毒载体整合的位置效应是影响外源基因表达的重要因素,慢病毒载体整合位点的研究是探索外源基因整合机制的手段之一.转基因整合位点研究的方法主要有5种:荧光原位杂交、个体基因组文库筛选法、反向PCR、接头PCR、锚定PCR.近来的研究后发现整合位点之间可能存在一些共同特征.

Research Approaches for Lentiviral Vector mediated Transgene Integration Site

Lentiviral vector is characterized with its stable integration into host genome and long-term expression in host cells, thus becoming one of the hotspots in development of gene therapeutic strategies and transgenic animal models.Integration position of lentiviral vector is a key factor affecting expression of transgene.To study the mechanism of transgene integration in host genome, first step is to clone the sequence of integration site.The approaches for this purpose include FISH, selection of individual genome library, inverse PCR, adaptor-PCR and anchored PCR.Different methods are conformed to different situations depending on how transgenes are produced.Recent studies have demonstrated that there appear some common characteristics among the integration-site sequences.