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Androgenetic haploid embryonic stem cells producelive transgenic mice.pdf

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Androgenetic haploid embryonic stem cells producelive transgenic mice

LETTER doi:10.1038/nature11435 Androgenetic haploid embryonic stem cells produce live transgenic mice Wei Li1,2*, Ling Shuai1,3*, Haifeng Wan1,2*, Mingzhu Dong1,2*, Meng Wang4, Lisi Sang1,2, Chunjing Feng1,2, Guan-Zheng Luo4, Tianda Li1,3, Xin Li1,3, Libin Wang1,2, Qin-Yuan Zheng1,2, Chao Sheng1,3, Hua-Jun Wu2,4, Zhonghua Liu3, Lei Liu1, Liu Wang1, Xiu-Jie Wang4, Xiao-Yang Zhao1 Qi Zhou1 Haploids and double haploids are important resources for study- ing recessive traits and have large impacts on crop breeding1, but natural haploids are rare in animals. Mammalian haploids are restricted to germline cells and are occasionally found in tumours with massive chromosome loss2,3. Recent success in establishing haploid embryonic stem (ES) cells in medaka fish4 and mice5,6 raised the possibility of using engineered mammalian haploid cells in genetic studies. However, the availability and functional characterization of mammalian haploid ES cells are still limited. Here we show that mouse androgenetic haploid ES (ahES) cell lines can be established by transferring sperm into an enucleated oocyte. The ahES cells maintain haploidy and stable growth over 30 passages, express pluripotent markers, possess the ability to differentiate into all three germ layers in vitro and in vivo, and contribute to germlines of chimaeraswhen injected into blastocysts. Although epigenetically distinct from sperm cells, the ahES cells can produce viable and fertile progenies after intracytoplasmic injection into mature oocytes. The oocyte-injection procedure can also produce viable transgenic mice from genetically engineered ahES cells. Our findings show the developmental pluripotency of androgenentic haploids and provide a new tool to quickly produce genetic models for recessive traits. They may also shed new light on assisted reproduction. Haploid stem cells offer an easy-to-manipulate genetic system and therefore hold great value for studies of recessive phenotypes. Recently, haploid ES cells h