| Title |
Generation of CRISPR/Cas9-mediated gene-targeted pigs via somatic cell nuclear transfer
|
|---|---|
| Published in |
Cellular and Molecular Life Sciences, October 2014
|
| DOI | 10.1007/s00018-014-1744-7 |
| Pubmed ID | |
| Authors |
Xiaoqing Zhou, Jige Xin, Nana Fan, Qingjian Zou, Jiao Huang, Zhen Ouyang, Yu Zhao, Bentian Zhao, Zhaoming Liu, Sisi Lai, Xiaoling Yi, Lin Guo, Miguel A. Esteban, Yangzhi Zeng, Huaqiang Yang, Liangxue Lai |
| Abstract |
The domestic pig has been widely used as an important large animal model. Precise and efficient genetic modification in pig provides a great promise in biomedical research. Recently, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system has been successfully used to produce many gene-targeted animals. However, these animals have been generated by co-injection of Cas9 mRNA and single-guide RNA (sgRNA) into one-cell stage embryos, which mostly resulted in mosaicism of the modification. One or two rounds of further breeding should be performed to obtain homozygotes with identical genotype and phenotype. To address this issue, gene-targeted somatic cells can be used as donor for somatic cell nuclear transfer (SCNT) to produce gene-targeted animals with single and identical mutations. In this study, we applied Cas9/sgRNAs to effectively direct gene editing in porcine fetal fibroblasts and then mutant cell colonies were used as donor to generate homozygous gene-targeted pigs through single round of SCNT. As a result, we successfully obtained 15 tyrosinase (TYR) biallelic mutant pigs and 20 PARK2 and PINK1 double-gene knockout (KO) pigs. They were all homozygous and no off-target mutagenesis was detected by comprehensive analysis. TYR (-/-) pigs showed typical albinism and the expression of parkin and PINK1 were depleted in PARK2 (-/-)/PINK1 (-/-) pigs. The results demonstrated that single- or double-gene targeted pigs can be effectively achieved by using the CRISPR/Cas9 system combined with SCNT without mosaic mutation and detectable off-target effects. This gene-editing system provides an efficient, rapid, and less costly manner to generate genetically modified pigs or other large animals. |
X Demographics
The data shown below were collected from the profiles of 3 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 67% |
| Russia | 1 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 67% |
| Scientists | 1 | 33% |
Mendeley readers
The data shown below were compiled from readership statistics for 160 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Japan | 1 | <1% |
| Spain | 1 | <1% |
| United States | 1 | <1% |
| Denmark | 1 | <1% |
| Unknown | 156 | 98% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 35 | 22% |
| Student > Ph. D. Student | 23 | 14% |
| Student > Master | 19 | 12% |
| Student > Bachelor | 19 | 12% |
| Student > Doctoral Student | 11 | 7% |
| Other | 18 | 11% |
| Unknown | 35 | 22% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 55 | 34% |
| Biochemistry, Genetics and Molecular Biology | 40 | 25% |
| Medicine and Dentistry | 4 | 3% |
| Veterinary Science and Veterinary Medicine | 3 | 2% |
| Immunology and Microbiology | 3 | 2% |
| Other | 13 | 8% |
| Unknown | 42 | 26% |
Attention Score in Context
This research output has an Altmetric Attention Score of 29. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 25 January 2017.
All research outputs
#1,216,563
of 23,794,258 outputs
Outputs from Cellular and Molecular Life Sciences
#99
of 4,151 outputs
Outputs of similar age
#13,943
of 255,295 outputs
Outputs of similar age from Cellular and Molecular Life Sciences
#3
of 51 outputs
Altmetric has tracked 23,794,258 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 94th percentile: it's in the top 10% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 4,151 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 6.0. This one has done particularly well, scoring higher than 99% of its peers.
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