دورية أكاديمية
Genome engineering of mammalian haploid embryonic stem cells using the Cas9/RNA system
العنوان: | Genome engineering of mammalian haploid embryonic stem cells using the Cas9/RNA system |
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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 |
تدمد: | 21678359 |
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DOI: | 10.7717/peerj.230 |