Gene expressing human artificial chromosome vectors: Advantages and challenges for gene therapy.
Gene expressing human artificial chromosome vectors: Advantages and challenges for gene therapy.
复制标题
基因表达人类人工染色体载体：基因治疗的优势和挑战。
DOI：
10.1016/j.yexcr.2020.111931
复制
发表时间：
2020
影响因子：
3.7
通讯作者：
Monaco,ZoiaL
中科院分区：
文献类型：
--
作者：
Moralli,Daniela;Monaco,ZoiaL
在20世纪80年代后期用酵母人工染色体构建基因组文库用于细胞中的基因分离和表达研究之后，人类人工染色体随后在20世纪90年代基于需要用于产生功能性人工染色体的着丝粒序列的相同形成原理而自然发展。在过去的二十年里，它们成为了解人类染色体结构和组织的有用研究工具，以及表达大基因和基因位点的重要载体和完全表达的调控区域。现在，它们正在被修饰和开发用于离体和体内的基因治疗。使用HAC载体的优点是它们保持自主性并表现为正常染色体。它们对于治疗研究是有吸引力的，而没有外源DNA整合到宿主染色体中的有害后果。与其他载体相比，HAC载体也是适应大序列（>100 kb）的唯一自主稳定载体。操纵这些载体以将基因有效递送到人细胞中的挑战仍在进行中，但我们已经在使用无辅助（HF）扩增子载体技术转移表达基因的HAC载体以在人细胞中产生从头HAC方面取得了进展。在单次处理中用两种HF扩增子同时感染后，成功实现了有效的多基因递送，并且输入DNA重组以形成从头HAC。潜在地，可以同时转导含有表达基因的HAC载体的几种扩增子，这将增加载体的基因负载能力，用于递送和研究在人细胞中的完全表达。
After the construction of genomic libraries with yeast artificial chromosomes in the late 1980's for gene isolation and expression studies in cells, human artificial chromosomes were then a natural development in the 1990's, based on the same principles of formation requiring centromeric sequences for generating functional artificial chromosomes. Over the past twenty years, they became a useful research tool for understanding human chromosome structure and organization, and important vectors for expression of large genes and gene loci and the regulatory regions for full expression. Now they are being modified and developed for gene therapy both ex vivo and in vivo. The advantages of using HAC vectors are that they remain autonomous and behave as a normal chromosome. They are attractive for therapy studies without the harmful consequences of integration of exogenous DNA into host chromosomes. HAC vectors are also the only autonomous stable vectors that accommodate large sequences (>100 kb) compared to other vectors. The challenges of manipulating these vectors for efficient delivery of genes into human cells is still ongoing, but we have made advances in transfer of gene expressing HAC vectors using the helper free (HF) amplicon vector technology for generating de novo HAC in human cells. Efficient multigene delivery was successfully achieved following simultaneous infection with two HF amplicons in a single treatment and the input DNA recombined to form a de novo HAC. Potentially several amplicons containing gene expressing HAC vectors could be transduced simultaneously which would increase the gene loading capacity of the vectors for delivery and studying full expression in human cells.