Genetic Engineering in the Pig: Gene Knockout and Alternative Techniques

ABSTRACT: Since endothelial cells (EC) are the major target cells during hyperacute rejection and are likely in delayed graft rejection, most of the genetic engineering of the xenotransplant donor is aimed at modifying their properties. Among the various strategies that are reviewed are the genotypic or phenotypic knockout of the αl,3Gal antigen, which is a major target of xenoantibodies and is also probably involved in innate cellular response. In addition, the success of the transgeny of complement regulatory proteins is well established. In vitro data from analyses of the mechanisms of endothelial cell activation also suggest that other molecules could be used to regulate apoptosis or thrombotic microenvironment or to minimize recipient T‐cell activation by inhibiting costimulatory proteins such as CD40 or B7. Alternative to usual knockout techniques (thus far not available in pigs, where no ES cells have been derived) will be presented.

[1]  E. Salvaris,et al.  Changes in cell surface glycosylation in alpha1,3-galactosyltransferase knockout and alpha1,2-fucosyltransferase transgenic mice. , 1997, Transplantation.

[2]  E. Field,et al.  Balancing the immune system for tolerance: a case for regulatory CD4 cells. , 1997, Transplantation.

[3]  F. Brook,et al.  The origin and efficient derivation of embryonic stem cells in the mouse. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Pardi,et al.  Human natural killer lymphocytes directly recognize evolutionarily conserved oligosaccharide ligands expressed by xenogeneic tissues. , 1997, Transplantation.

[5]  D. Cramer,et al.  Human serum reactivity to porcine endothelial cells after antisense-mediated down-regulation of GpIIIa expression. , 1997, Transplantation.

[6]  I. Wilmut,et al.  "Viable Offspring Derived from Fetal and Adult Mammalian Cells" (1997), by Ian Wilmut et al. , 2014 .

[7]  F. Bach,et al.  Inhibition of bovine endothelial cell activation in vitro by regulated expression of a transdominant inhibitor of NF-kappa B. , 1997, The Journal of clinical investigation.

[8]  D. Candinas,et al.  Accommodation of vascularized xenografts: Expression of “protective genes” by donor endothelial cells in a host Th2 cytokine environment , 1997, Nature Medicine.

[9]  J. Platt,et al.  Transgenic pigs expressing human CD59 and decay-accelerating factor produce an intrinsic barrier to complement-mediated damage. , 1997, Transplantation.

[10]  A. Gutiérrez-Adán,et al.  Isolation of pluripotent stem cells from cultured porcine primordial germ cells. , 1997, Biology of reproduction.

[11]  A. Skalka,et al.  Intracellular expression of single-chain variable fragments to inhibit early stages of the viral life cycle by targeting human immunodeficiency virus type 1 integrase , 1996, Journal of Virology.

[12]  M. Wright,et al.  The use of intracellular single-chain antibody fragments to inhibit specifically the expression of cell surface molecules. , 1996, Journal of immunological methods.

[13]  D. Alexander,et al.  Protein tyrosine phosphatases in T-cell development, apoptosis and signalling. , 1996, Immunology today.

[14]  A. Dalmasso,et al.  Assessment of hyperacute rejection in a rat-to-primate cardiac xenograft model. , 1996, Transplantation.

[15]  J. Renard,et al.  Cloning of domestic species , 1996 .

[16]  C. Ferran,et al.  Expression of a truncated form of the human p55 TNF-receptor in bovine aortic endothelial cells renders them resistant to human TNF. , 1996, Transplantation Proceedings.

[17]  José Alberola-lla,et al.  The use of dominant-negative mutations to elucidate signal transduction pathways in lymphocytes. , 1996, Current opinion in immunology.

[18]  I. Wilmut,et al.  Sheep cloned by nuclear transfer from a cultured cell line , 1996, Nature.

[19]  Inhibition of endothelial cell activation by adenovirus-mediated expression of I kappa B alpha, an inhibitor of the transcription factor NF-kappa B , 1996, The Journal of experimental medicine.

[20]  E. Setter,et al.  Enzymatic remodelling of the carbohydrate surface of a xenogenic cell substantially reduces human antibody binding and complement-mediated cytolysis , 1995, Nature Medicine.

[21]  K. Stuhlmeier,et al.  Barriers to xenotransplantation , 1995, Nature Medicine.

[22]  M. Gerritsen,et al.  Transfection of the human heme oxygenase gene into rabbit coronary microvessel endothelial cells: protective effect against heme and hemoglobin toxicity. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[23]  David Baltimore,et al.  Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-κB , 1995, Nature.

[24]  M. Reitz,et al.  Intracellular expression of antibody fragments directed against HIV reverse transcriptase prevents HIV infection in vitro , 1995, Nature Medicine.

[25]  T. Waldmann,et al.  Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the alpha subunit of the receptor. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[26]  C. Baldari,et al.  Inhibition of CD4/p56lck signaling by a dominant negative mutant of the Shc adaptor protein. , 1995, Oncogene.

[27]  J. Markmann,et al.  Survival Of Mhc‐Deficient Mouse Heterotopic Cardiac Allografts , 1995, Transplantation.

[28]  L. Matis,et al.  Expression of a functional human complement inhibitor in a transgenic pig as a model for the prevention of xenogeneic hyperacute organ rejection. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  N. Hynes,et al.  Autocrine inhibition of the epidermal growth factor receptor by intracellular expression of a single-chain antibody. , 1994, Biochemical and biophysical research communications.

[30]  O. Bagasra,et al.  Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Cattaneo,et al.  Intracellular Immunization with Cytosolic Recombinant Antibodies , 1994, Bio/Technology.

[32]  M. Wheeler,et al.  Development and validation of swine embryonic stem cells: a review. , 1994, Reproduction, fertility, and development.

[33]  M. Korner,et al.  NF‐kappa B activates the HIV promoter in neurons. , 1993, The EMBO journal.

[34]  S. Chen,et al.  Design, intracellular expression, and activity of a human anti-human immunodeficiency virus type 1 gp120 single-chain antibody. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Philip M. Murphy,et al.  Molecular mimicry and the generation of host defense protein diversity , 1993, Cell.

[36]  R. Gibbs,et al.  Fidelity of targeted recombination in human fibroblasts and murine embryonic stem cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[37]  B. Wasylyk,et al.  Transformation suppressor activity of a Jun transcription factor lacking its activation domain , 1991, Nature.

[38]  R. Prather,et al.  Nuclear transfer in mammalian embryos. , 1990, International review of cytology.

[39]  H. Eibel,et al.  Elimination of CD8+ thymocytes in transgenic mice expressing an anti‐Lyt2.2 immunoglobulin heavy chain gene. , 1989, EMBO Journal.

[40]  J. Lunney,et al.  Localization of class II MHC antigens on porcine renal vascular endothelium. , 1984, Transplantation.

[41]  R. J. Mullen,et al.  Relationship of genotype and degree of chimerism in coat color to sex ratios and gametogenesis in chimeric mice. , 1971, The Journal of experimental zoology.