Forward and Reverse Genetics through Derivation of Haploid Mouse Embryonic Stem Cells

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 ES cell 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 the first knock-out 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 bioweapon. Our results open the possibility to combine 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.

A haploid genetics toolbox for Arabidopsis thaliana

“…Haploid genetics has long provided an invaluable tool for basic genetic research in both prokaryote and eukaryote microbial systems [17][18][19] and has recently been extended to mammalian systems through the generation of haploid cell lines 20,21 . However, although Arabidopsis remains a premiere model system for basic research in higher plants, haploid genetics has been largely inaccessible to the Arabidopsis research community.…”

Generation of human haploid embryonic stem cells from parthenogenetic embryos obtained by microsurgical removal of male pronucleus

“…Haploid embryonic stem cells (haESCs) have been recently generated from parthenogenetic (PG) or androgenetic (AG) blastocysts of different mammals, including mouse, rat and monkey, enabling genetic screening at both cellular and organism levels [1][2][3][4][5][6]. However, whether haESCs can be generated from human remains unknown.…”

Derivation and application of pluripotent stem cells for regenerative medicine

“…Since the 1970s, extensive efforts have been made to generate haploid embryos in the mouse (Tarkowski and Rossant, 1976), but it was not until 2011 that the mouse parthenogenetic haploid ESCs (phESCs) were derived from parthenogenetic haploid embryos via fluorescence-activated cell sorting of haploid cells (Elling et al, 2011;Leeb and Wutz, 2011). Moreover, mouse androgenetic haploid ESCs (ahESCs) which can function as sperm were established in 2012 Yang et al, 2012).…”