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Androgenetic haploid embryonic stem cells produce live transgenic mice

Abstract

Haploids and double haploids are important resources for studying 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 chimaeras when 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.

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Figure 1: Generation of androgenetic haploid ES cells.
Figure 2: The pluripotency of the ahES cells.
Figure 3: Generation of ICAI offspring and transgenic mice.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

CGH data and gene-expression data are deposited at the Gene Expression Omnibus under accession numbers GSE39390 and GSE39391, respectively.

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Acknowledgements

We thank all members of the Group of Reproductive Engineering for discussion and help. This study was supported by a grant from the National Science Foundation of China 90919060 (to Q.Z.) and the China National Basic Research Program 2012CBA01300 (to Q.Z.), and a grant from the Strategic Priority Research Program of the Chinese Academy of Sciences XDA01020100 (to Q.Z.). We thank Fluidigm Corporation for their support in the utilization of BioMark HD system. We thank Eppendorf and Leica for supporting the facility.

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Q.Z. and X.-Y.Z. designed the experiments;; W.L., L.S., H.W., M.D., M.W., L.S., C.F., T.L., X.L., L.W., Q.-Y.Z. and C.S. performed experiments; Q.Z, X.-Y.Z, X.-J.W., G.-Z.L., W.L., H.-J.W. and L.L. analysed data; Q.Z., X.-Y.Z., Z.L., and L.W..supervised experiments; W.L., L.S., H.W. and M.D. contributed to part of the Methods. Q.Z., X.-Y.Z., X.-J.W. and W.L. wrote the paper.

Corresponding authors

Correspondence to Xiao-Yang Zhao or Qi Zhou.

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The authors declare no competing financial interests.

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This file contains Supplementary Figures 1-15, Supplementary Tables 1-6 and an additional reference. (PDF 1628 kb)

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Li, W., Shuai, L., Wan, H. et al. Androgenetic haploid embryonic stem cells produce live transgenic mice. Nature 490, 407–411 (2012). https://doi.org/10.1038/nature11435

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