Highly Efficient Transfer of Chromosomes to a Broad Range of Target Cells Using Chinese Hamster Ovary Cells Expressing Murine Leukemia Virus-Derived Envelope Proteins.
Highly Efficient Transfer of Chromosomes to a Broad Range of Target Cells Using Chinese Hamster Ovary Cells Expressing Murine Leukemia Virus-Derived Envelope Proteins.
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DOI：
10.1371/journal.pone.0157187
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Hara T
中科院分区：
文献类型：
--
作者：
Suzuki T;Kazuki Y;Oshimura M;Hara T
微细胞介导的染色体转移（MMCT）是将供体细胞的染色体导入受体细胞的重要步骤。MMCT不仅允许对特定染色体进行遗传/表观遗传分析，还允许利用人类和小鼠人工染色体（HAC/MAC）作为基因递送载体。尽管对基因组规模分析的科学需求不断增加，但现有方法的低转移效率阻碍了染色体工程技术的应用。在这里，我们开发了一种高效的染色体转移方法，称为retro-MMCT，这是基于中国仓鼠卵巢细胞表达来自嗜亲性或嗜亲性小鼠白血病病毒的包膜蛋白。使用这种方法，我们将MAC转移到NIH 3 T3细胞，其效率比使用常规MMCT方法获得的效率高26.5倍。Retro-MMCT适用于各种受体细胞，包括胚胎干细胞。此外，retro-MMCT能够将MAC有效地转移至来自人、猴、小鼠、大鼠和兔的受体细胞。这些结果证明了逆转录MMCT用于将染色体有效转移到各种类型的靶细胞的效用。
Microcell-mediated chromosome transfer (MMCT) is an essential step for introducing chromosomes from donor cells to recipient cells. MMCT allows not only for genetic/epigenetic analysis of specific chromosomes, but also for utilization of human and mouse artificial chromosomes (HACs/MACs) as gene delivery vectors. Although the scientific demand for genome scale analyses is increasing, the poor transfer efficiency of the current method has hampered the application of chromosome engineering technology. Here, we developed a highly efficient chromosome transfer method, called retro-MMCT, which is based on Chinese hamster ovary cells expressing envelope proteins derived from ecotropic or amphotropic murine leukemia viruses. Using this method, we transferred MACs to NIH3T3 cells with 26.5 times greater efficiency than that obtained using the conventional MMCT method. Retro-MMCT was applicable to a variety of recipient cells, including embryonic stem cells. Moreover, retro-MMCT enabled efficient transfer of MAC to recipient cells derived from humans, monkeys, mice, rats, and rabbits. These results demonstrate the utility of retro-MMCT for the efficient transfer of chromosomes to various types of target cell.