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

被引：226

作者：

Elling, Ulrich ^{[1
]}

Taubenschmid, Jasmin ^{[1
]}

Wirnsberger, Gerald ^{[1
]}

O'Malley, Ronan ^{[2
]}

Demers, Simon-Pierre ^{[3
]}

Vanhaelen, Quentin ^{[3
]}

Shukalyuk, Andrey I. ^{[4
]}

Schmauss, Gerald ^{[1
]}

Schramek, Daniel ^{[1
]}

Schnuetgen, Frank ^{[5
]}

von Melchner, Harald ^{[5
]}

Ecker, Joseph R. ^{[2
,7
]}

Stanford, William L. ^{[3
,4
,6
]}

Zuber, Johannes ^{[8
]}

Stark, Alexander ^{[8
]}

Penninger, Josef M. ^{[1
]}

机构：

[1] Austrian Acad Sci, Inst Mol Biotechnol, IMBA, A-1030 Vienna, Austria

[2] Salk Inst Biol Studies, Genom Anal Lab, La Jolla, CA 92037 USA

[3] Ottawa Hosp Res Inst, Sprott Ctr Stem Cell Res, Ottawa, ON K1H 8L6, Canada

[4] Univ Toronto, Toronto, ON M5G 1L5, Canada

[5] Goethe Univ Frankfurt, Sch Med, Dept Mol Hematol, D-60590 Frankfurt, Germany

[6] Inst Syst Biol, Seattle, WA USA

[7] Howard Hughes Med Inst, Seattle, WA USA

[8] Inst Mol Pathol, A-1030 Vienna, Austria

来源：

CELL STEM CELL | 2011年 / 9卷 / 06期

基金：

欧洲研究理事会; 奥地利科学基金会;

关键词：

SELF-RENEWAL; TRANSCRIPTIONAL NETWORKS; EXPRESSION; DIFFERENTIATION; PLURIPOTENCY; COMMITMENT; SCREENS; NUMBER; GENES; LINE;

D O I：

10.1016/j.stem.2011.10.012

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：563 / 574

页数：12

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