دورية أكاديمية
The CRISPR/Cas9 Minipig—A Transgenic Minipig to Produce Specific Mutations in Designated Tissues
| العنوان: | The CRISPR/Cas9 Minipig—A Transgenic Minipig to Produce Specific Mutations in Designated Tissues |
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| المؤلفون: | Berthelsen, Martin Fogtmann, Riedel, Maria, Cai, Huiqiang, Skaarup, Søren H., Alstrup, Aage K.O., Dagnæs-Hansen, Frederik, Luo, Yonglun, Jensen, Uffe B., Hager, Henrik, Liu, Ying, Callesen, Henrik, Vendelbo, Mikkel H., Jakobsen, Jannik E., Thomsen, Martin Kristian |
| المصدر: | Berthelsen , M F , Riedel , M , Cai , H , Skaarup , S H , Alstrup , A K O , Dagnæs-Hansen , F , Luo , Y , Jensen , U B , Hager , H , Liu , Y , Callesen , H , Vendelbo , M H , Jakobsen , J E & Thomsen , M K 2021 , ' The CRISPR/Cas9 Minipig—A Transgenic Minipig to Produce Specific Mutations in Designated Tissues ' , Cancers , vol. 13 , no. 12 , 3024 . https://doi.org/10.3390/cancers13123024Test |
| سنة النشر: | 2021 |
| المجموعة: | Aarhus University: Research |
| مصطلحات موضوعية: | Animal models, Cancer, CRISPR, Göttingen minipigs, Handmade cloning, KRAS, Lung cancer, Porcine model, STK11, TP53 |
| الوصف: | The generation of large transgenic animals is impeded by complex cloning, long maturation and gastrulation times. An introduction of multiple gene alterations increases the complexity. We have cloned a transgenic Cas9 minipig to introduce multiple mutations by CRISPR in somatic cells. Transgenic Cas9 pigs were generated by somatic cell nuclear transfer and were backcrossed to Göttingen Minipigs for two generations. Cas9 expression was controlled by FlpO-mediated recombination and was visualized by translation from red to yellow fluorescent protein. In vitro analyses in primary fibroblasts, keratinocytes and lung epithelial cells confirmed the genetic alterations executed by the viral delivery of single guide RNAs (sgRNA) to the target cells. Moreover, multiple gene alterations could be introduced simultaneously in a cell by viral delivery of sgRNAs. Cells with loss of TP53, PTEN and gain-of-function mutation in KRAS G12D showed increased proliferation, confirming a transformation of the primary cells. An in vivo activation of Cas9 expression could be induced by viral delivery to the skin. Overall, we have generated a minipig with conditional expression of Cas9, where multiple gene alterations can be introduced to somatic cells by viral delivery of sgRNA. The development of a transgenic Cas9 minipig facilitates the creation of complex pre-clinical models for cancer research. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| العلاقة: | https://pure.au.dk/portal/en/publications/f1f00d41-0209-4a56-92ce-a2bd8dd947acTest |
| DOI: | 10.3390/cancers13123024 |
| الإتاحة: | https://doi.org/10.3390/cancers13123024Test https://pure.au.dk/portal/en/publications/f1f00d41-0209-4a56-92ce-a2bd8dd947acTest http://www.scopus.com/inward/record.url?scp=85107897352&partnerID=8YFLogxKTest |
| حقوق: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.F41E376F |
| قاعدة البيانات: | BASE |
| DOI: | 10.3390/cancers13123024 |
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