Highly Efficient Transfer of Chromosomes to a Broad Range of Target Cells Using Chinese Hamster Ovary Cells Expressing Murine Leukemia Virus-Derived Envelope Proteins.
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.
Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
Teruhiko
http://orcid.org/0000-0001-8162-8809
T
Suzuki
Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, Tottori, Japan.|Chromosome Engineering Research Center, Tottori University, Tottori, Japan.
Yasuhiro
Y
Kazuki
Chromosome Engineering Research Center, Tottori University, Tottori, Japan.
Mitsuo
M
Oshimura
Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
Takahiko
T
Hara
D000818:Animals; D016466:CHO Cells; D022201:Chromosomes, Artificial / Q000235:genetics*; D006224:Cricetinae; D003412:Cricetulus; D015686:Gene Products, env / Q000235:genetics / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D011817:Rabbits; D051381:Rats
D015686:Gene Products, env