Institute of Experimental Hematology, Hannover Medical School, D-30625 Hannover, Germany.
Hum Gene Ther. 2013 Feb;24(2):132-42. doi: 10.1089/hum.2012.229.
Over the past decades, lentiviral vectors have evolved as a benchmark tool for stable gene transfer into cells with a high replicative potential. Their relatively flexible genome and ability to transduce many forms of nondividing cells, combined with the potential for cell-specific pseudotyping, provides a rich resource for numerous applications in experimental platforms and therapeutic settings. Here, we give an overview of important biosafety features of lentiviral vectors, with detailed discussion of (i) the principles of the lentiviral split-genome design used for the construction of packaging cells; (ii) the relevance of modifications introduced into the lentiviral long terminal repeat (deletion of enhancer/promoter sequences and introduction of insulators); (iii) the basic features of mRNA processing, including the Rev/Rev-responsive element (RRE) interaction and the modifications of the 3' untranslated region of lentiviral vectors with various post-transcriptional regulatory elements affecting transcriptional termination, polyadenylation, and differentiation-specific degradation of mRNA; and (iv) the characteristic integration pattern with the associated risk of transcriptional interference with cellular genes. We conclude with considerations regarding the importance of cell targeting via envelope modifications. Along this course, we address canonical biosafety issues encountered with any type of viral vector: the risks of shedding, mobilization, germline transmission, immunogenicity, and insertional mutagenesis.
在过去的几十年中，慢病毒载体已发展成为将基因稳定转入具有高复制潜能的细胞的基准工具。其相对灵活的基因组和转导多种非分裂细胞的能力，加上细胞特异性假型化的潜力，为实验平台和治疗环境中的许多应用提供了丰富的资源。在这里，我们概述了慢病毒载体的重要生物安全特性，并详细讨论了（i）用于构建包装细胞的慢病毒分裂基因组设计的原理；（ii）引入慢病毒长末端重复序列中的修饰（增强子/启动子序列的缺失和绝缘子的引入）；（iii）mRNA 加工的基本特征，包括 Rev/Rev 反应元件（RRE）相互作用以及各种转录后调控元件对慢病毒载体 3'非翻译区的修饰，影响转录终止、多聚腺苷酸化和 mRNA 的分化特异性降解；和（iv）特征性整合模式以及与细胞基因转录干扰相关的风险。最后，我们考虑了通过包膜修饰进行细胞靶向的重要性。在此过程中，我们解决了任何类型的病毒载体都会遇到的典型生物安全问题：脱落、移动、种系传播、免疫原性和插入突变的风险。
