Park Frank
Department of Medicine, Kidney Disease Center, Medical College of Wisconsin, Wauwatosa, Wisconsin 53226, USA.
Physiol Genomics. 2007 Oct 22;31(2):159-73. doi: 10.1152/physiolgenomics.00069.2007. Epub 2007 Aug 7.
Lentiviral vectors have become a promising new tool for the establishment of transgenic animals and the manipulation of the mammalian genome. While conventional microinjection-based methods for transgenesis have been successful in generating small and large transgenic animals, their relatively low transgenic efficiency has opened the door for alternative approaches, including lentiviral vectors. Lentiviral vectors are an appealing tool for transgenesis in part because of their ability to incorporate into genomic DNA with high efficiency, especially in cells that are not actively dividing. Lentiviral vector-mediated transgene expression can also be maintained for long periods of time. Recent studies have documented high efficiencies for lentiviral transgenesis, even in animal species and strains, such as NOD/scid and C57Bl/6 mouse, that are very difficult to manipulate using the standard transgenic techniques. These advantages of the lentiviral vector system have broadened its use as a gene therapy vector to additional applications that include transgenesis and knockdown functional genetics. This review will address the components of the lentiviral vector system and recent successes in lentiviral transgenesis using both male- and female-derived pluripotent cells. The advantages and disadvantages of lentiviral transgenesis vs. other approaches to produce transgenic animals will be compared with regard to efficiency, the ability to promote persistent transgene expression, and the time necessary to generate a sufficient number of animals for phenotyping.
慢病毒载体已成为用于建立转基因动物和操纵哺乳动物基因组的一种有前景的新工具。虽然基于传统显微注射的转基因方法已成功产生了大小各异的转基因动物,但其相对较低的转基因效率为包括慢病毒载体在内的替代方法打开了大门。慢病毒载体是一种有吸引力的转基因工具,部分原因在于其能够高效整合到基因组DNA中,尤其是在不活跃分裂的细胞中。慢病毒载体介导的转基因表达也能长时间维持。最近的研究记录了慢病毒转基因的高效率,即使在使用标准转基因技术很难操作的动物物种和品系中,如NOD/scid和C57Bl/6小鼠。慢病毒载体系统的这些优势已将其作为基因治疗载体的用途扩展到包括转基因和敲低功能遗传学在内的其他应用中。本综述将探讨慢病毒载体系统的组成部分以及使用雄性和雌性来源的多能细胞进行慢病毒转基因的最新成功案例。将在效率、促进转基因持续表达的能力以及产生足够数量用于表型分析的动物所需的时间方面,比较慢病毒转基因与其他生产转基因动物方法的优缺点。