Highly Efficient Transfer of Chromosomes to a Broad Range of Target Cells Using Chinese Hamster Ovary Cells Expressing Murine Leukemia Virus-Derived Envelope Proteins

被引：32

作者：

Suzuki, Teruhiko ^{[1
]}

Kazuki, Yasuhiro ^{[2
,3
]}

Oshimura, Mitsuo ^{[3
]}

Hara, Takahiko ^{[1
]}

机构：

[1] Tokyo Metropolitan Inst Med Sci, Stem Cell Project, Tokyo 113, Japan

[2] Tottori Univ, Dept Biomed Sci, Inst Regenerat Med & Biofunct, Grad Sch Med Sci, Tottori 680, Japan

[3] Tottori Univ, Chromosome Engn Res Ctr, Tottori 680, Japan

来源：

PLOS ONE | 2016年 / 11卷 / 06期

基金：

日本学术振兴会;

关键词：

HUMAN ARTIFICIAL CHROMOSOMES; FUSION;

D O I：

10.1371/journal.pone.0157187

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：11

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