The production of multi-transgenic pigs: update and perspectives for xenotransplantation

2016 ◽  
Vol 25 (3) ◽  
pp. 361-374 ◽  
Author(s):  
Heiner Niemann ◽  
Bjoern Petersen
Keyword(s):  
Transgenic Pigs

scholarly journals Trichostatin A-Assisted Epigenomic Modulation Affects the Expression Profiles of Not Only Recombinant Human α1,2-Fucosyltransferase and α-Galactosidase A Enzymes But Also Galα1→3Gal Epitopes in Porcine Bi-Transgenic Adult Cutaneous Fibroblast Cells

2021 ◽  
Vol 22 (3) ◽  
pp. 1386
Author(s):  
Jerzy Wiater ◽  
Marcin Samiec ◽  
Maria Skrzyszowska ◽  
Daniel Lipiński
Keyword(s):  
Genetic Resource ◽  
Ex Vivo ◽  
Trichostatin A ◽  
Expression Profiles ◽  
Fibroblast Cells ◽  
Control Groups ◽  
Transgenic Pigs ◽  
Cell Tissue ◽  
Donor Cells ◽  
Lower Expression

This study was conducted to explore whether trichostatin A-assisted epigenomic modulation (TSA-EM) can affect the expression of not only recombinant human α1,2-fucosyltransferase (rhα1,2-FT) and α-galactosidase A (rhα-Gal A) immune system enzymes but also Galα1→3Gal epitopes in ex vivo proliferating adult cutaneous fibroblast cells (ACFCs) derived from hFUT2×hGLA bi-transgenic pigs that had been produced for the needs of future xenotransplantation efforts. The ACFC lines were treated with 50 nM TSA for 24 h and then the expression profiles of rhα1,2-FT and rhα-Gal A enzymes were analyzed by Western blot and immunofluorescence. The expression profiles of the Galα1→3Gal epitope were determined by lectin blotting and lectin fluorescence. The ACFCs derived from non-transgenic (nTG) pigs were served as the negative (TSA−) and positive (TSA+) control groups. For both hFUT2×hGLA and nTG samples, the expression levels of α1,2-FT and α-Gal A proteins in TSA+ cells were more than twofold higher in comparison to TSA− cells. Moreover, a much lower expression of the Galα1→3Gal epitopes was shown in TSA− hFUT2×hGLA cells as compared to the TSA− nTG group. Interestingly, the levels of Galα1→3Gal expression in TSA-treated hFUT2×hGLA and nTG ACFCs were significantly higher than those noticed for their TSA-untreated counterparts. Summing up, ex vivo protection of effectively selected bi-transgenic ACFC lines, in which TSA-dependent epigenetic transformation triggered the enhancements in reprogrammability and subsequent expression of hFUT2 and hGLA transgenes and their corresponding transcripts, allows for cryopreservation of nuclear donor cells, nuclear-transferred female gametes, and resultant porcine cloned embryos. The latter can be used as a cryogenically conserved genetic resource of biological materials suitable for generation of bi-transgenic cloned offspring in pigs that is targeted at biomedical research in the field of cell/tissue xenotransplantation.

Introduction of a porcine growth hormone fusion gene into transgenic pigs promotes growth

1988 ◽  
Vol 90 (2) ◽  
pp. 295-300
Author(s):  
P.D. Vize ◽  
A.E. Michalska ◽  
R. Ashman ◽  
B. Lloyd ◽  
B.A. Stone ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Fusion Gene ◽  
Transgenic Animals ◽  
Growth Hormone Gene ◽  
Gene Sequences ◽  
Transgenic Pigs ◽  
Gene Construct ◽  
Metallothionein Promoter

Six transgenic pigs have been produced by microinjecting a human metallothionein promoter/porcine growth hormone gene construct into the pronuclei of fertilized eggs which were transferred to synchronized recipient sows. The resulting transgenic animals contained between 0.5 and 15 copies of the gene construct per cell, and at least one of the animals expressed the introduced gene and grew at an increased rate compared to both transgenic and non-transgenic littermates. Some of the transgenic animals that did not appear to grow at increased rates were found to contain rearranged gene sequences. Two of the transgenic pigs have been shown to pass on the introduced genes to their offspring.

scholarly journals Determination of the Absolute Number of Transgene Copies in CMVFUT Transgenic Pigs

2012 ◽  
Vol 12 (3) ◽  
pp. 349-356 ◽  
Author(s):  
Daniel Lipiński ◽  
Joanna Zeyland ◽  
Andrzej Pławski ◽  
Ryszard Słomski
Keyword(s):  
Copy Number ◽  
Transgenic Animals ◽  
Promoter Sequence ◽  
Absolute Number ◽  
Transgenic Pigs ◽  
Cmv Promoter ◽  
Genetic Construct ◽  
Coding Sequence ◽  
The Absolute

Determination of the Absolute Number of Transgene Copies in CMVFUT Transgenic PigsThe aim of this research was to determine the number of transgene copies in the DNA of transgenic pigs. The copy number of the transgene was analysed in the transgenic animals with introduced pCMVFUT genetic construct containing a coding sequence of human H transferase under a control of CMV promoter. The copy number of the transgene that had integrated with the genome of the transgenic animals was analysed by qPCR with SYBR Green dye, which enabled nonspecific double-stranded DNA detection. CMVFT-2F and CMVFT-2R primers were used to amplify a 149 bp fragment of DNA. Forward primer had a sequence complementary to a promoter sequence and reverse primer to a coding sequence of H transferase. The copy number of the transgene in the examined samples was established by plotting the CT values obtained on a standard curve, which had been set by the usage of the CT values for the successive standard dilutions with known copy number (1.438-1.431 copies). As a standard we used pCMVFut genetic construct hydrolyzed with Not I restriction enzyme to a linear form. The real-time PCR results helped to establish the range of 3 - 4 as the number of the transgene copies that had integrated to the swine genome.

scholarly journals Production of Recombinant Human Von Willebrand Factor in the Milk of Transgenic Pigs

2009 ◽  
Vol 55 (5) ◽  
pp. 484-490 ◽  
Author(s):  
Hyun-Gi LEE ◽  
Hwi-Cheul LEE ◽  
Sung Woo KIM ◽  
Poongyeon LEE ◽  
Hak-Jae CHUNG ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Transgenic Pigs ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals Generation of cloned transgenic pigs rich in omega-3 fatty acids

2006 ◽  
Vol 24 (4) ◽  
pp. 435-436 ◽  
Author(s):  
Liangxue Lai ◽  
Jing X Kang ◽  
Rongfeng Li ◽  
Jingdong Wang ◽  
William T Witt ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Omega 3 Fatty Acids ◽  
Omega 3 ◽  
Transgenic Pigs

Cytolytic assessment of hyperacute rejection and production of nuclear transfer embryos using hCD46-transgenic porcine embryonic germ cells

Zygote ◽  
2009 ◽  
Vol 17 (2) ◽  
pp. 101-108 ◽  
Author(s):  
Ji Young Won ◽  
Kwang Sung Ahn ◽  
Alice M. Sorrell ◽  
Susa Shin ◽  
Soon Young Heo ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Regulatory Protein ◽  
Blastocyst Stage ◽  
Hyperacute Rejection ◽  
Complement Regulatory Protein ◽  
Transgenic Pigs ◽  
Embryonic Germ Cells ◽  
Transgenic Embryos ◽  
Stage Analysis ◽  
Human Xenotransplantation

SummaryHuman complement regulatory protein hCD46 may reduce the hyperacute rejection (HAR) in pig-to-human xenotransplantation. In this study, an hCD46 gene was introduced into porcine embryonic germ (EG) cells. Treatment of human serum did not affect the survival of hCD46-transgenic EG cells, whereas the treatment significantly reduced the survival of non-transgenic EG cells (p < 0.01). The transgenic EG cells presumably capable of alleviating HAR were transferred into enucleated oocytes. Among 235 reconstituted oocytes, 35 (14.9%) developed to the blastocyst stage. Analysis of individual embryos indicated that 80.0% (28/35) of embryos contained the transgene hCD46. The result of the present study demonstrates resistance of hCD46-transgenic EG cells against HAR, and the usefulness of the transgenic approach may be predicted by this cytolytic assessment prior to actual production of transgenic pigs. Subsequently performed EG cell nuclear transfer gave rise to hCD46-transgenic embryos. Further study on the transfer of these embryos to recipients may produce hCD46-transgenic pigs.

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