Division of Pulmonary and Critical Care Medicine, Northwestern University, Chicago, IL, USA.
Gene Ther. 2010 Apr;17(4):439-47. doi: 10.1038/gt.2010.31. Epub 2010 Mar 4.
The nuclear envelope represents a key barrier to successful nonviral transfection and gene therapy both in vitro and in vivo. Although the main purpose of the nuclear envelope is to partition the cell to maintain cytoplasmic components in the cytoplasm and nuclear components, most notably genomic DNA, in the nucleus, this function poses a problem for transfections in which exogenous DNA is delivered into the cytoplasm. After delivery to the cytoplasm, nucleic acids rapidly become complexed with cellular proteins that mediate interactions with the cellular machinery for trafficking. Thus, it is these proteins that, in essence, control the nuclear import of DNA, and we must also understand their activities in cells. In this review, we will discuss the principles of nuclear import of proteins and DNA-protein complexes, as well as the various approaches that investigators have used to improve nuclear targeting of plasmids. These approaches include complexation of plasmids with peptides, native and engineered proteins, ligands and polymers, as well as the inclusion of transcription factor-binding sites for general and cell-specific delivery.
nonviral gene transfermid R:plasmidmid R:nuclear pore complexmid R:importinmid R:nuclear localization signalmid R:karyopherin.
核膜是成功进行非病毒转染和基因治疗的关键障碍，无论是在体外还是体内。尽管核膜的主要目的是分隔细胞，以保持细胞质成分在细胞质中，核成分，最显著的是基因组 DNA，在细胞核中，这个功能对转染提出了一个问题，其中外源性 DNA 被递送到细胞质中。在递送到细胞质后，核酸迅速与细胞蛋白复合，这些蛋白介导与细胞机器的运输相互作用。因此，正是这些蛋白质实质上控制着 DNA 的核内输入，我们也必须了解它们在细胞中的活性。在这篇综述中，我们将讨论蛋白质和 DNA-蛋白质复合物的核内输入的原理，以及研究人员为提高质粒的核靶向性而采用的各种方法。这些方法包括将质粒与肽、天然和工程蛋白、配体和聚合物复合，以及包括转录因子结合位点，以实现一般和细胞特异性的传递。
非病毒基因转移；质粒；核孔复合体；导入蛋白；核定位信号；核输出蛋白。
