Transfer of a Mouse Artificial Chromosome into Spermatogonial Stem Cells Generates Transchromosomic Mice.
Transfer of a Mouse Artificial Chromosome into Spermatogonial Stem Cells Generates Transchromosomic Mice.
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DOI：
10.1016/j.stemcr.2017.08.012
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发表时间：
2017-10-10
影响因子：
5.9
通讯作者：
Kazuki Y
中科院分区：
文献类型：
--
作者：
Shinohara T;Kazuki K;Ogonuki N;Morimoto H;Matoba S;Hiramatsu K;Honma K;Suzuki T;Hara T;Ogura A;Oshimura M;Kanatsu-Shinohara M;Kazuki Y
将百万碱基大小的DNA片段引入生殖系一直是一项艰巨的任务。虽然微细胞介导的染色体转移到小鼠胚胎干细胞(ESCs)可以产生跨染色体的小鼠，但ESCs的核型不稳定，通过嵌合体的形成进行生殖系传播是不可靠的。由于精原干细胞(SSCs)是生殖系中唯一的干细胞，它们是生殖系修饰的一个有吸引力的靶点。在这里，我们报告了成功地将小鼠人工染色体(MAC)转移到小鼠生殖系干细胞(GSCs)，培养的精原细胞富含SSCs。与染色体数目有显著差异的ESCs相比，转MAC的GSCs更稳定地保持了宿主的核型和MAC。此外，转移了MAC的GSCs在显微注射到不育受者的生精小管后产生了跨染色体小鼠。成功地将MSC转移到GSCs克服了与ESC介导的生殖系传播相关的问题，并为生殖系修饰提供了新的可能性。Rtro-MMCT方法允许将小鼠人工染色体转移到GSCs中GSCs维护的外源染色体比ESCs更稳定跨染色体小鼠是生殖细胞移植后从GSCs产生的小鼠不同于ESCs，跨染色体小鼠直接出生在F1代在本文中，Kazuki及其同事证明精原干细胞能够比ESCs更稳定地维持微染色体载体，ESCs在长期培养过程中经常会经历核型异常和微小染色体的丢失。由于跨染色体小鼠可以在F1代产生，这种方法克服了以前使用ESCs的方法相关的问题，并为生殖系操作提供了新的可能性。
The introduction of megabase-sized large DNA fragments into the germline has been a difficult task. Although microcell-mediated chromosome transfer into mouse embryonic stem cells (ESCs) allows the production of transchromosomic mice, ESCs have unstable karyotypes and germline transmission is unreliable by chimera formation. As spermatogonial stem cells (SSCs) are the only stem cells in the germline, they represent an attractive target for germline modification. Here, we report successful transfer of a mouse artificial chromosome (MAC) into mouse germline stem cells (GSCs), cultured spermatogonia enriched for SSCs. MAC-transferred GSCs maintained the host karyotype and MAC more stably than ESCs, which have significant variation in chromosome number. Moreover, MAC-transferred GSCs produced transchromosomic mice following microinjection into the seminiferous tubules of infertile recipients. Successful transfer of MACs to GSCs overcomes the problems associated with ESC-mediated germline transmission and provides new possibilities in germline modification. Retro-MMCT method allows transfer of a mouse artificial chromosome into GSCs GSCs maintained exogenous chromosomes more stably than ESCs Transchromosomic mice were born from GSCs following germ cell transplantation Unlike ESCs, transchromosomic mice were born directly in F1 generation In this article, Kazuki and colleagues show that spermatogonial stem cells can maintain a minichromosome vector more stably than ESCs, which often undergo karyotype abnormalities and loss of minichromosomes during long-term culture. Because transchromosomic mice can be produced in F1 generation, this method overcomes problems associated with previous methods using ESCs and provides new possibilities for germline manipulation.
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DOI：
10.1073/pnas.0508780102
发表时间：
2005-11-29
影响因子：
11.1
作者：
Hamra, FK;Chapman, KM;Garbers, DL
通讯作者：
Garbers, DL
DOI：
10.1073/pnas.0605755103
发表时间：
2006-08-29
影响因子：
11.1
作者：
Ogonuki, Narumi;Mochida, Keiji;Ogura, Atsuo
通讯作者：
Ogura, Atsuo
影响因子：
3.6
作者：
Kanatsu-Shinohara, Mito;Ogonuki, Narumi;Shinohara, Takashi
通讯作者：
Shinohara, Takashi
影响因子：
3.6
作者：
Morimoto, Hiroko;Lee, Jiyoung;Shinohara, Takashi
通讯作者：
Shinohara, Takashi
影响因子：
2.6
作者：
Oshimura, Mitsuo;Uno, Narumi;Kazuki, Yasuhiro;Katoh, Motonobu;Inoue, Toshiaki
通讯作者：
Inoue, Toshiaki