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Cell Stem Cell. 2011 Dec 2;9(6):563-74. doi: 10.1016/j.stem.2011.10.012.

Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.

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  • 1IMBA, Institute of Molecular Biotechnology of the Austrian Academy of Sciences, 1030 Vienna, Austria.

Abstract

All somatic mammalian cells carry two copies of chromosomes (diploidy), whereas organisms with a single copy of their genome, such as yeast, provide a basis for recessive genetics. Here we report the generation of haploid mouse ESC lines from parthenogenetic embryos. These cells carry 20 chromosomes, express stem cell markers, and develop into all germ layers in vitro and in vivo. We also developed a reversible mutagenesis protocol that allows saturated genetic recessive screens and results in homozygous alleles. This system allowed us to generate a knockout cell line for the microRNA processing enzyme Drosha. In a forward genetic screen, we identified Gpr107 as a molecule essential for killing by ricin, a toxin being used as a bioweapon. Our results open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale.

Copyright © 2011 Elsevier Inc. All rights reserved.

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PMID:
22136931
[PubMed - indexed for MEDLINE]
PMCID:
PMC4008724
Free PMC Article
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