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Genome engineering of mammalian haploid embryonic stem cells using the Cas9/RNA system

التفاصيل البيبلوغرافية
العنوان: Genome engineering of mammalian haploid embryonic stem cells using the Cas9/RNA system
المؤلفون: Takuro Horii, Sumiyo Morita, Mika Kimura, Ryouhei Kobayashi, Daiki Tamura, Ryou-u Takahashi, Hironobu Kimura, Isao Suetake, Hirokazu Ohata, Koji Okamoto, Shoji Tajima, Takahiro Ochiya, Yumiko Abe, Izuho Hatada
المصدر: PeerJ, Vol 1, p e230 (2013)
بيانات النشر: PeerJ Inc., 2013.
سنة النشر: 2013
المجموعة: LCC:Medicine
LCC:Biology (General)
مصطلحات موضوعية: CRISPR/Cas, Haploid, Embryonic stem cells, Genome engineering, Tet1, Tet2, Medicine, Biology (General), QH301-705.5
الوصف: Haploid embryonic stem cells (ESCs) are useful for studying mammalian genes because disruption of only one allele can cause loss-of-function phenotypes. Here, we report the use of haploid ESCs and the CRISPR RNA-guided Cas9 nuclease gene-targeting system to manipulate mammalian genes. Co-transfection of haploid ESCs with vectors expressing Cas9 nuclease and single-guide RNAs (sgRNAs) targeting Tet1, Tet2, and Tet3 resulted in the complete disruption of all three genes and caused a loss-of-function phenotype with high efficiency (50%). Co-transfection of cells with vectors expressing Cas9 and sgRNAs targeting two loci on the same chromosome resulted in the creation of a large chromosomal deletion and a large inversion. Thus, the use of the CRISPR system in combination with haploid ESCs provides a powerful platform to manipulate the mammalian genome.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2167-8359
Relation: https://peerj.com/articles/230.pdf; https://peerj.com/articles/230/; https://doaj.org/toc/2167-8359
DOI: 10.7717/peerj.230
URL الوصول: https://doaj.org/article/afb4ac3fd52f4d40abac7a2467ea58f0
رقم الأكسشن: edsdoj.fb4ac3fd52f4d40abac7a2467ea58f0
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:21678359
DOI:10.7717/peerj.230