Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system
Using gene editing technology based on CRISPR/Cas9n system and somatic cell nuclear transfer technology, we constructed a plasmid of two sgRNAs and Cas9 nickase, and green fluorescent protein (GFP) in the same vector followed by transient transfection of porcine fibroblasts. By flow cytometry, the c...
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| Format: | Article |
| Language: | English |
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Zhejiang University Press
2018-03-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.03.013 |
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| author | WANG Shaohua ZHAO Panpan LIU Tong DING Biao LUO Lei CAO Zubing ZHANG Yunhai ZHANG Kun |
| author_facet | WANG Shaohua ZHAO Panpan LIU Tong DING Biao LUO Lei CAO Zubing ZHANG Yunhai ZHANG Kun |
| author_sort | WANG Shaohua |
| collection | DOAJ |
| description | Using gene editing technology based on CRISPR/Cas9n system and somatic cell nuclear transfer technology, we constructed a plasmid of two sgRNAs and Cas9 nickase, and green fluorescent protein (GFP) in the same vector followed by transient transfection of porcine fibroblasts. By flow cytometry, the cells expressing GFP were selected, and the GFP positive cells were cultured to single cells. The CD163 edited clones were screened through polymerase chain reaction (PCR) and DNA sequencing, and the positive rate was as high as 90% (18/20). Somatic cell nuclear transfer and embryo transfer were carried out using cells with edited CD163, and two CD163 gene-edited cloned offsprings were obtained. |
| format | Article |
| id | doaj-art-e4f89c4531944a78a14ab6df3cd9907a |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2018-03-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-e4f89c4531944a78a14ab6df3cd9907a2025-08-20T03:32:11ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552018-03-014415716110.3785/j.issn.1008-9209.2018.03.01310089209Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n systemWANG ShaohuaZHAO PanpanLIU TongDING BiaoLUO LeiCAO ZubingZHANG YunhaiZHANG KunUsing gene editing technology based on CRISPR/Cas9n system and somatic cell nuclear transfer technology, we constructed a plasmid of two sgRNAs and Cas9 nickase, and green fluorescent protein (GFP) in the same vector followed by transient transfection of porcine fibroblasts. By flow cytometry, the cells expressing GFP were selected, and the GFP positive cells were cultured to single cells. The CD163 edited clones were screened through polymerase chain reaction (PCR) and DNA sequencing, and the positive rate was as high as 90% (18/20). Somatic cell nuclear transfer and embryo transfer were carried out using cells with edited CD163, and two CD163 gene-edited cloned offsprings were obtained.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.03.013CRISPR/Cas9n systemgene editingblue ear disease<italic>CD163</italic> gene |
| spellingShingle | WANG Shaohua ZHAO Panpan LIU Tong DING Biao LUO Lei CAO Zubing ZHANG Yunhai ZHANG Kun Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system 浙江大学学报. 农业与生命科学版 CRISPR/Cas9n system gene editing blue ear disease <italic>CD163</italic> gene |
| title | Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system |
| title_full | Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system |
| title_fullStr | Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system |
| title_full_unstemmed | Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system |
| title_short | Production of porcine reproductive and respiratory syndrome virus (PRRSV)-resistant genomeedited cloned pigs using CRISPR/Cas9n system |
| title_sort | production of porcine reproductive and respiratory syndrome virus prrsv resistant genomeedited cloned pigs using crispr cas9n system |
| topic | CRISPR/Cas9n system gene editing blue ear disease <italic>CD163</italic> gene |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.03.013 |
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