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An EBV-Based Plasmid Can Replicate and Maintain in Stem Cells

机译:基于EBV的质粒可以在干细胞中复制和维持

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摘要

Viral vectors have a wide range of applications in biology, particularly in gene therapy. Based on their integration capacity, viral vectors are classified as either integrating or non-integrating vectors. Although integrating vectors, such as lentivectors, have the ability to direct prolonged expression of exogenous genes, manipulation of the host genome is an inappropriate feature of these gene delivery tools. Non-integrating vectors, such as episomal replicating plasmids, can replicate and persist in host cells for long periods without any chromosomal interruption. These advantages made them good tools for gene induction purposes in gene therapy and basic studies. Due to the necessity of gene induction in stem cells for study of mammalian development and targeted differentiation, the use of integrating vectors for prolonged expression of genes of interest has been developed. Application of replicating plasmids can overcome some drawbacks associated with integrating vectors, although replication and maintenance of these plasmids can differ between cell types. Previously, it has been shown that such plasmids can be maintained in human embryonic stem cells for more than one month, but the rate of the plasmid replication during the host cell cycle has not been elucidated. In the present study, we showed that an EBV-based plasmid can replicate simultaneously with host in pluripotent and multipotent human and mouse stem cells and can be sustained for long time periods in dividing cells. (C) 2015 American Institute of Chemical Engineers
机译:病毒载体在生物学特别是基因治疗中具有广泛的应用。基于它们的整合能力,病毒载体被分类为整合或非整合载体。尽管整合的载体,例如慢病毒载体,具有指导外源基因延长表达的能力,但是宿主基因组的操纵是这些基因递送工具的不合适的特征。非整合型载体,例如附加型复制质粒,可以在宿主细胞中复制并持续很长时间,而不会中断任何染色体。这些优势使它们成为基因治疗和基础研究中用于基因诱导目的的良好工具。由于在干细胞中进行基因诱导以研究哺乳动物的发育和靶向分化的必要性,已经开发了整合载体用于延长目的基因表达的用途。复制质粒的应用可以克服与整合载体相关的一些缺点,尽管这些质粒的复制和维持在细胞类型之间可能有所不同。以前,已经表明这种质粒可以在人胚胎干细胞中维持超过一个月,但是尚未阐明在宿主细胞周期中质粒复制的速率。在本研究中,我们表明基于EBV的质粒可以在多能和多能人和小鼠干细胞中与宿主同时复制,并且可以在分裂细胞中长期维持。 (C)2015美国化学工程师学会

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