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Open AccessJournal ArticleDOI

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

TLDR
The generation of haploid mouse ESC lines from parthenogenetic embryos is reported, opening 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.
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This article is published in Cell Stem Cell.The article was published on 2011-12-02 and is currently open access. It has received 214 citations till now. The article focuses on the topics: Cellular differentiation & Somatic cell.

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Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide RNA library

TL;DR: The results demonstrate the potential for efficient loss-of-function screening using the CRISPR-Cas9 system and identify 27 known and 4 previously unknown genes implicated in these phenotypes.
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Target Identification for Small Bioactive Molecules: Finding the Needle in the Haystack

TL;DR: Current methods for target identification of small molecules are summarized, primarily for a chemistry audience but also the biological community, for example, the chemist or biologist attempting to identify the target of a given bioactive compound.
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Thiol-linked alkylation of RNA to assess expression dynamics.

TL;DR: Thiol(SH)-linked alkylation for the metabolic sequencing of RNA (SLAM seq), an orthogonal-chemistry-based RNA sequencing technology that detects 4-thiouridine incorporation in RNA species at single-nucleotide resolution, facilitates the dissection of fundamental mechanisms that control gene expression in an accessible, cost-effective and scalable manner.
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The ‘obligate diploid’ Candida albicans forms mating-competent haploids

TL;DR: It is demonstrated that C. albicans has a viable haploid state that can be derived from diploid cells under in vitro and in vivo conditions, and that seems to arise through a concerted chromosome loss mechanism.
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Megabase-scale deletion using CRISPR/Cas9 to generate a fully haploid human cell line

TL;DR: In this paper, the authors employed a CRISPR/Cas9-based genome engineering strategy to excise this sizeable chromosomal fragment and to efficiently and reproducibly derive clones that retain their haploid state.
References
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Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.

TL;DR: Induction of pluripotent stem cells from mouse embryonic or adult fibroblasts by introducing four factors, Oct3/4, Sox2, c-Myc, and Klf4, under ES cell culture conditions is demonstrated and iPS cells, designated iPS, exhibit the morphology and growth properties of ES cells and express ES cell marker genes.
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The nuclear RNase III Drosha initiates microRNA processing

TL;DR: The two RNase III proteins, Drosha and Dicer, may collaborate in the stepwise processing of miRNAs, and have key roles in miRNA-mediated gene regulation in processes such as development and differentiation.
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A System for Stable Expression of Short Interfering RNAs in Mammalian Cells

TL;DR: It is shown that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes.
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Mutations affecting segment number and polarity in Drosophila

TL;DR: The phenotypes of the mutant embryos indicate that the process of segmentation involves at least three levels of spatial organization: the entire egg as developmental unit, a repeat unit with the length of two segments, and the individual segment.
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Functional expression cloning of nanog, a pluripotency sustaining factor in embryonic stem cells

TL;DR: These findings establish a central role for Nanog in the transcription factor hierarchy that defines ES cell identity and confirm that Cytokine dependence, multilineage differentiation, and embryo colonization capacity are fully restored upon transgene excision.
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