Generation by somatic cell nuclear transfer of GGTA1 knockout pigs expressing soluble human TNFRI-Fc and human HO-1

Geon A. Kim; Eun Mi Lee; Bumrae Cho; Zahid Alam; Su Jin Kim; Sanghoon Lee; Hyun Ju Oh; Jong Ik Hwang; Curie Ahn; Byeong Chun Lee

doi:10.1007/s11248-018-0103-0

Generation by somatic cell nuclear transfer of GGTA1 knockout pigs expressing soluble human TNFRI-Fc and human HO-1

Geon A. Kim, Eun Mi Lee, Bumrae Cho, Zahid Alam, Su Jin Kim, Sanghoon Lee, Hyun Ju Oh, Jong Ik Hwang, Curie Ahn, Byeong Chun Lee

Department of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

Herein, we successfully generated transgenic pigs expressing both soluble human tumor necrosis factor receptor I IgG ₁ -Fc (shTNFRI-Fc) and human hemagglutinin (HA)-tagged-human heme oxygenase-1 (hHO-1) without Gal epitope. Healthy cloned pigs were produced by somatic cell nuclear transfer (SCNT) using the genetically modified cells. The genetic disruption of the GGTA1 genes and absence of expression of BS-IB4 lectin in tail-derived fibroblast of the SCNT-generated piglets were successfully confirmed. The expression of shTNFRI-Fc and HAhHO-1 was fully identified with protective effect against oxidative stress and apoptosis stimulation. Antibody-mediated complement-dependent cytotoxicity assay for examining the immuno-reactivity of transgenically derived pig cells showed that pigs lacking GGTA1 with the expression of double genes reduce the humoral barrier to xenotransplantation, more than pigs simply expressing double genes and the wild type. Through this approach, rapid production of a pig strain deficient in various genes may be expected to be applicable for xenotransplantation research without extensive breeding protocols.

Original language	English
Pages (from-to)	91-102
Number of pages	12
Journal	Transgenic Research
Volume	28
Issue number	1
DOIs	https://doi.org/10.1007/s11248-018-0103-0
Publication status	Published - 2019 Feb 1

Keywords

Alpha1,3-galactosyltransferase gene
Genetically engineered pigs
Soluble human TNFRI-Fc and human HO-1
Somatic cell nuclear transfer
Xenotransplantation

ASJC Scopus subject areas

Biotechnology
Animal Science and Zoology
Agronomy and Crop Science
Genetics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/s11248-018-0103-0

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@article{cd0e79b6bd384f07af893a27857b27c1,

title = "Generation by somatic cell nuclear transfer of GGTA1 knockout pigs expressing soluble human TNFRI-Fc and human HO-1",

abstract = " Herein, we successfully generated transgenic pigs expressing both soluble human tumor necrosis factor receptor I IgG 1 -Fc (shTNFRI-Fc) and human hemagglutinin (HA)-tagged-human heme oxygenase-1 (hHO-1) without Gal epitope. Healthy cloned pigs were produced by somatic cell nuclear transfer (SCNT) using the genetically modified cells. The genetic disruption of the GGTA1 genes and absence of expression of BS-IB4 lectin in tail-derived fibroblast of the SCNT-generated piglets were successfully confirmed. The expression of shTNFRI-Fc and HAhHO-1 was fully identified with protective effect against oxidative stress and apoptosis stimulation. Antibody-mediated complement-dependent cytotoxicity assay for examining the immuno-reactivity of transgenically derived pig cells showed that pigs lacking GGTA1 with the expression of double genes reduce the humoral barrier to xenotransplantation, more than pigs simply expressing double genes and the wild type. Through this approach, rapid production of a pig strain deficient in various genes may be expected to be applicable for xenotransplantation research without extensive breeding protocols. ",

keywords = "Alpha1,3-galactosyltransferase gene, Genetically engineered pigs, Soluble human TNFRI-Fc and human HO-1, Somatic cell nuclear transfer, Xenotransplantation",

author = "Kim, {Geon A.} and Lee, {Eun Mi} and Bumrae Cho and Zahid Alam and Kim, {Su Jin} and Sanghoon Lee and Oh, {Hyun Ju} and Hwang, {Jong Ik} and Curie Ahn and Lee, {Byeong Chun}",

note = "Funding Information: Acknowledgements We thank Dr. Peter Cowan for generously providing the GTKO porcine endothelial cells. This study was supported by the Ministry of Trade, Industry & Energy (#10048948), National Research Foundation (#2015R1C1A2A01054373; 2016M3A9B6903410), Research Institute for Veterinary Science, Natural Balance and the BK21 plus program. This research was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean Government, MSIP (2014M3A9D3034034). These supporters had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscripts. Publisher Copyright: {\textcopyright} 2018, Springer Nature Switzerland AG.",

year = "2019",

month = feb,

day = "1",

doi = "10.1007/s11248-018-0103-0",

language = "English",

volume = "28",

pages = "91--102",

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T1 - Generation by somatic cell nuclear transfer of GGTA1 knockout pigs expressing soluble human TNFRI-Fc and human HO-1

AU - Kim, Geon A.

AU - Lee, Eun Mi

AU - Cho, Bumrae

AU - Alam, Zahid

AU - Kim, Su Jin

AU - Lee, Sanghoon

AU - Oh, Hyun Ju

AU - Hwang, Jong Ik

AU - Ahn, Curie

AU - Lee, Byeong Chun

N1 - Funding Information: Acknowledgements We thank Dr. Peter Cowan for generously providing the GTKO porcine endothelial cells. This study was supported by the Ministry of Trade, Industry & Energy (#10048948), National Research Foundation (#2015R1C1A2A01054373; 2016M3A9B6903410), Research Institute for Veterinary Science, Natural Balance and the BK21 plus program. This research was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean Government, MSIP (2014M3A9D3034034). These supporters had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscripts. Publisher Copyright: © 2018, Springer Nature Switzerland AG.

PY - 2019/2/1

Y1 - 2019/2/1

N2 - Herein, we successfully generated transgenic pigs expressing both soluble human tumor necrosis factor receptor I IgG 1 -Fc (shTNFRI-Fc) and human hemagglutinin (HA)-tagged-human heme oxygenase-1 (hHO-1) without Gal epitope. Healthy cloned pigs were produced by somatic cell nuclear transfer (SCNT) using the genetically modified cells. The genetic disruption of the GGTA1 genes and absence of expression of BS-IB4 lectin in tail-derived fibroblast of the SCNT-generated piglets were successfully confirmed. The expression of shTNFRI-Fc and HAhHO-1 was fully identified with protective effect against oxidative stress and apoptosis stimulation. Antibody-mediated complement-dependent cytotoxicity assay for examining the immuno-reactivity of transgenically derived pig cells showed that pigs lacking GGTA1 with the expression of double genes reduce the humoral barrier to xenotransplantation, more than pigs simply expressing double genes and the wild type. Through this approach, rapid production of a pig strain deficient in various genes may be expected to be applicable for xenotransplantation research without extensive breeding protocols.

AB - Herein, we successfully generated transgenic pigs expressing both soluble human tumor necrosis factor receptor I IgG 1 -Fc (shTNFRI-Fc) and human hemagglutinin (HA)-tagged-human heme oxygenase-1 (hHO-1) without Gal epitope. Healthy cloned pigs were produced by somatic cell nuclear transfer (SCNT) using the genetically modified cells. The genetic disruption of the GGTA1 genes and absence of expression of BS-IB4 lectin in tail-derived fibroblast of the SCNT-generated piglets were successfully confirmed. The expression of shTNFRI-Fc and HAhHO-1 was fully identified with protective effect against oxidative stress and apoptosis stimulation. Antibody-mediated complement-dependent cytotoxicity assay for examining the immuno-reactivity of transgenically derived pig cells showed that pigs lacking GGTA1 with the expression of double genes reduce the humoral barrier to xenotransplantation, more than pigs simply expressing double genes and the wild type. Through this approach, rapid production of a pig strain deficient in various genes may be expected to be applicable for xenotransplantation research without extensive breeding protocols.

KW - Alpha1,3-galactosyltransferase gene

KW - Genetically engineered pigs

KW - Soluble human TNFRI-Fc and human HO-1

KW - Somatic cell nuclear transfer

KW - Xenotransplantation

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DO - 10.1007/s11248-018-0103-0

M3 - Article

C2 - 30552552

AN - SCOPUS:85058397029

SN - 0962-8819

VL - 28

SP - 91

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JO - Transgenic Research

JF - Transgenic Research

IS - 1

ER -

Generation by somatic cell nuclear transfer of GGTA1 knockout pigs expressing soluble human TNFRI-Fc and human HO-1

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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