Earticle

초록

영어

To avoid hyperacute rejection of xenografts, α1,3-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of α1,3-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that α2,3-sialyltransferase (α2,3ST) and α2,6-sialyltransferase (α2,6ST) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of α-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of α2,6-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Construction of α1,3-galactosyltransferase Gene KO Vector
  PCR Analysis of Targeted Cells
  Production of Heterozygote GalT KO Pigs
  Sample Preparation and Protein Determination
  RNA Isolation and Real-time RT-PCR
  Assay of Glycosyltransferase Activity
  Lectin Blotting
  Analysis of Sialic Acid Contents
  Statistical Analysis
 RESULTS
  Production of Heterozygote GalT KO Gnotobiotic Miniature Pigs
  Heterozygote GalT KO Pig-Derived Liver Displays Higher Non-Gal Xenoantigen
 DISCUSSION
 REFERENCES

키워드

저자

• Gyesik Min [ Department of Pharmaceutical Engineering, Gyeongnam National University of Science & Technology ]
• Jong-Yi Park [ Department of Animal Biotechnology, Konkuk University ] Corresponding author

참고문헌

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간행물

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• 간행물명

  Reproductive & developmental biology

• 간기

  계간

• pISSN

  1738-2432

• eISSN

  2288-0151

• 수록기간

  1977~2018

• 십진분류

  KDC 527 DDC 636