Laboratory of Molecular Pharmacology, NCI, NIH, Bethesda, MD, USA.
Cell Mol Life Sci. 2013 Apr;70(7):1135-48. doi: 10.1007/s00018-012-1113-3. Epub 2012 Aug 21.
Since their description in the late 1990s, human artificial chromosomes (HACs) carrying a functional kinetochore were considered as a promising system for gene delivery and expression with a potential to overcome many problems caused by the use of viral-based gene transfer systems. Indeed, HACs avoid the limited cloning capacity, lack of copy number control and insertional mutagenesis due to integration into host chromosomes that plague viral vectors. Nevertheless, until recently, HACs have not been widely recognized because of uncertainties of their structure and the absence of a unique gene acceptor site. The situation changed a few years ago after engineering of HACs with a single loxP gene adopter site and a defined structure. In this review, we summarize recent progress made in HAC technology and concentrate on details of two of the most advanced HACs, 21HAC generated by truncation of human chromosome 21 and alphoid(tetO)-HAC generated de novo using a synthetic tetO-alphoid DNA array. Multiple potential applications of the HAC vectors are discussed, specifically the unique features of two of the most advanced HAC cloning systems.
自 20 世纪 90 年代末描述以来，携带功能性着丝粒的人源人工染色体（HAC）被认为是一种很有前途的基因传递和表达系统，有可能克服许多由病毒基因转移系统引起的问题。事实上，HAC 避免了由于整合到宿主染色体而导致的病毒载体所存在的克隆容量有限、缺乏拷贝数控制和插入突变等问题。然而，直到最近，由于其结构的不确定性和缺乏独特的基因接受位点，HAC 并没有得到广泛认可。几年前，通过在 HAC 上工程改造一个单一的 loxP 基因接受位点和一个明确的结构，这种情况发生了改变。在这篇综述中，我们总结了 HAC 技术的最新进展，并集中讨论了两种最先进的 HAC 的细节，即通过截断人类 21 号染色体产生的 21HAC 和使用合成的 tetO-alphoid DNA 阵列从头生成的 alphoid(tetO)-HAC。讨论了 HAC 载体的多种潜在应用，特别是两种最先进的 HAC 克隆系统的独特特征。
