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Cas9-expressing chickens and pigs as resources for genome editing in livestock [Agricultural Sciences]
Proceedings of the National Academy of Sciences of the United States of America ( IF 9.4 ) Pub Date : 2021-03-09 , DOI: 10.1073/pnas.2022562118
Beate Rieblinger 1 , Hicham Sid 2 , Denise Duda 2 , Tarik Bozoglu 3, 4 , Romina Klinger 2 , Antonina Schlickenrieder 2 , Kamila Lengyel 2 , Krzysztof Flisikowski 1 , Tatiana Flisikowska 1 , Nina Simm 1 , Alessandro Grodziecki 1 , Carolin Perleberg 1 , Andrea Bähr 3, 4 , Lucie Carrier 5, 6 , Mayuko Kurome 7, 8 , Valeri Zakhartchenko 7, 8 , Barbara Kessler 7, 8 , Eckhard Wolf 7, 8 , Lutz Kettler 9 , Harald Luksch 9 , Ibrahim T Hagag 10 , Daniel Wise 11 , Jim Kaufman 11, 12 , Benedikt B Kaufer 13 , Christian Kupatt 4, 14 , Angelika Schnieke 15 , Benjamin Schusser 16
Affiliation  

Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.



中文翻译:


表达 Cas9 的鸡和猪作为牲畜基因组编辑的资源 [农业科学]



转基因动物继续为健康和疾病的分子基础提供重要的见解。研究主要集中在转基因小鼠上,尽管猪等其他物种的生理机能更接近人类。此外,与系统发育较远的物种(例如鸡)的跨物种比较为基本的生物学和生物医学过程提供了强有力的见解。 CRISPR-Cas9 是适用于跨物种的最通用的遗传方法之一。在这里,我们报告了在所有器官中组成型表达 Cas9 的转基因鸡和猪的产生。这些动物健康且能生育。 Cas9 的功能在两个物种中都得到了证实,针对多种不同的靶基因、多种细胞类型以及体内的情况,通过淋巴细胞和发育中的大脑中的靶基因破坏,以及通过精确切除心脏中的 12.7 kb DNA 片段。 Cas9转基因动物将为农业和转化生物医学研究的体内基因组编辑提供强大的资源,并将促进反向遗传学以及跨物种比较。

更新日期:2021-03-04
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