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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Lucas-hahn, Andrea Hauschild, Janet Queisser, Anna-lisa Carnwath, Joseph W. Meng, Xiangdong Petersen, Bjoern Schwinzer, Reinhard Cost, Gregory J. Gregory, Philip D. Niemann, Heiner Santiago, Yolanda Zhang, Lei |
| Description | Author Affiliation: Hauschild J ( Friedrich-Loeffler-Institut, Mariensee, 31535 Neustadt am Rübenberge, Germany.); |
| Abstract | Zinc-finger nucleases (ZFNs) are powerful tools for producing gene knockouts (KOs) with high efficiency. Whereas ZFN-mediated gene disruption has been demonstrated in laboratory animals such as mice, rats, and fruit flies, ZFNs have not been used to disrupt an endogenous gene in any large domestic species. Here we used ZFNs to induce a biallelic knockout of the porcine 1,3-galactosyltransferase (GGTA1) gene. Primary porcine fibroblasts were treated with ZFNs designed against the region coding for the catalytic core of GGTA1, resulting in biallelic knockout of â ¼1% of ZFN-treated cells. A galactose (Gal) epitope counter-selected population of these cells was used in somatic cell nuclear transfer (SCNT). Of the resulting six fetuses, all completely lacked Gal epitopes and were phenotypically indistinguishable from the starting donor cell population, illustrating that ZFN-mediated genetic modification did not interfere with the cloning process. Neither off-target cleavage events nor integration of the ZFN-coding plasmid was detected. The GGTA1-KO phenotype was confirmed by a complement lysis assay that demonstrated protection of GGTA1-KO fibroblasts relative to wild-type cells. Cells from GGTA1-KO fetuses and pooled, transfected cells were used to produce live offspring via SCNT. This study reports the production of cloned pigs carrying a biallelic ZFN-induced knockout of an endogenous gene. These findings open a unique avenue toward the creation of gene KO pigs, which could benefit both agriculture and biomedicine. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 29 |
| Volume Number | 108 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2011-07-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Cloning, Organism Deoxyribonucleases Metabolism Galactosyltransferases Genetics Gene Knockout Techniques Sus Scrofa Animals DNA Primers Fibroblasts Flow Cytometry Molecular Sequence Data Nuclear Transfer Techniques Sequence Analysis, DNA Transplantation, Heterologous Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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