Transgenic expression of the human A20 gene in cloned pigs provides protection against apoptotic and inflammatory stimuli

Oropeza, Marianne; Petersen, Björn; Carnwath, Joseph Wallace; Lucas‐Hahn, Andrea; Lemme, Erika; Hassel, Petra; Herrmann, Doris; Barg-Kues, Brigitte; Holler, Stephanie; Queisser, A. L.; Schwinzer, Reinhard; Hinkel, Rabea; Kupatt, Christian; Niemann, Heiner

doi:10.1111/j.1399-3089.2009.00556.x

Cited by 122 publications

(83 citation statements)

References 58 publications

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“…The presence of three degraded fetuses was likely due to selection in the uterus against false or incomplete nuclear reprogramming (21). On the basis of our previous extensive experience with the somatic cloning of transgenic porcine cells (2,22,23) we conclude that the relatively high rate of embryonic loss in this experiment is likely due to the SCNT procedure and not ZFN treatment. Pigs carrying a GGTA1-KO are known to suffer from low birth weights, a low cloning efficiency, and health problems (14).…”

Section: Discussionmentioning

confidence: 80%

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Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases

Hauschild

Petersen

Santiago

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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334

251

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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 α1,3-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 ∼1% 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.

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Section: Discussionmentioning

confidence: 80%

“…Correspondingly, all fetuses showed a significantly decreased susceptibility to complement-mediated lysis comparable to the HR-derived GGTA1-KO cells described previously (23,29).…”

Section: Discussionmentioning

confidence: 87%

Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases

Hauschild

Petersen

Santiago

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

334

251

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“…Without the human complement regulator genes on the surface of PERV, antiGal-alpha-1,3-Gal antibodies prevent virus infection (207,208). In addition to the complement regulators, genes involved in adaptive immune responses and coagulation, e.g., the HLA-E/human beta2-microglobulin (338), the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) (160), the tumor necrosis factor alpha-induced protein 3 (A20) (240), thrombomodulin (251), and human heme oxygenase-1 (hHO-1) (252), have also been expressed in transgenic pigs.…”

Section: Use Of Transgenic Pigs To Overcome Immunological Rejectionmentioning

confidence: 99%

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

2012

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SUMMARY Xenotransplantation may be a solution to overcome the shortage of organs for the treatment of patients with organ failure, but it may be associated with the transmission of porcine microorganisms and the development of xenozoonoses. Whereas most microorganisms may be eliminated by pathogen-free breeding of the donor animals, porcine endogenous retroviruses (PERVs) cannot be eliminated, since these are integrated into the genomes of all pigs. Human-tropic PERV-A and -B are present in all pigs and are able to infect human cells. Infection of ecotropic PERV-C is limited to pig cells. PERVs may adapt to host cells by varying the number of LTR-binding transcription factor binding sites. Like all retroviruses, they may induce tumors and/or immunodeficiencies. To date, all experimental, preclinical, and clinical xenotransplantations using pig cells, tissues, and organs have not shown transmission of PERV. Highly sensitive and specific methods have been developed to analyze the PERV status of donor pigs and to monitor recipients for PERV infection. Strategies have been developed to prevent PERV transmission, including selection of PERV-C-negative, low-producer pigs, generation of an effective vaccine, selection of effective antiretrovirals, and generation of animals transgenic for a PERV-specific short hairpin RNA inhibiting PERV expression by RNA interference.

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“…In mice, expression of human endothelial protein C receptor (hEPCR) on islets improved graft survival and function, reduced inflammation and coagulation, and allowed diabetes correction using less islets than required when wild-type mice were used as donors [37]. Antiapoptotic and anti-inflammatory molecules heme oxygenase-1 (HO-1) and A20 were shown to be efficient in preserving transgenic porcine endothelial cells against TNF-α-induced apoptosis [38,39]. In islets, expression of HO-1 prolonged pig-to-mouse graft survival and decreased immune cell infiltration and islet cell apoptosis [40].…”

Section: Modification Of Donor Pigs To Mitigate the Immunementioning

confidence: 99%

Gene Editing, Gene Therapy, and Cell Xenotransplantation: Cell Transplantation Across Species

Mourad

Gianello

2017

Curr Transpl Rep

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Purpose of Review Cell xenotransplantation has the potential to provide a safe, ethically acceptable, unlimited source for cell replacement therapies. This review focuses on genetic modification strategies aimed to overcome remaining hurdles standing in the way of clinical porcine islet transplantation and to develop neural cell xenotransplantation. Recent Findings In addition to previously described genetic modifications aimed to mitigate hyperacute rejection, instant blood-mediated inflammatory reaction, and cell-mediated rejection, new data showing the possibility of increasing porcine islet insulin secretion by transgenesis is an interesting addition to the array of genetically modified pigs available for xenotransplantation. Moreover, combining multiple modifications is possible today thanks to new, improved genomic editing tools. Summary Genetic modification of large animals, pigs in particular, has come a long way during the last decade. These modifications can help minimize immunological and physiological incompatibilities between porcine and human cells, thus allowing for better tolerance and function of xenocells.

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Transgenic expression of the human A20 gene in cloned pigs provides protection against apoptotic and inflammatory stimuli

Cited by 122 publications

References 58 publications

Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases

Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases

Infection Barriers to Successful Xenotransplantation Focusing on Porcine Endogenous Retroviruses

Gene Editing, Gene Therapy, and Cell Xenotransplantation: Cell Transplantation Across Species

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