alpha1,3-Galactosyltransferase-gene knockout in cattle using a single targeting vector with loxP sequences and cre-expressing adenovirus

Transplantation
Yutaka SendaiYoshito Aoyagi

Abstract

Gene targeting in large animals has the potential to be useful in medicine as well as in agriculture. Previously, we reported the first successful targeting of the bovine alpha1,3-galactosyltransferase (alpha1,3GT) gene and establishment of a heterozygous knockout cell line. In this report, we generated both heterozygous and homozygous knockout bovine cell lines, and alpha1,3GT-gene knockout cattle. alpha1,3GT gene-disruption was accomplished using primary fetal fibroblasts with a single targeting vector, a promoter-less positive selection vector containing IRES (internal ribosome entry site)-antibiotic-resistance gene (neo) cassette and loxP sequences. At each step in establishing heterozygous and homozygous knockout cell lines, the antibiotic-resistance gene cassette in the targeted allele was removed by a Cre-loxP recombination system that utilizes an adenovirus with transient Cre recombinase expression. A nuclear transfer was performed using alpha1,3GT fetal fibroblasts, and one alpha1,3GT knockout calf was generated but died shortly after birth (day 287). Necropsy revealed normal morphology in all organs. The calf weighed 22.3 kg at birth and this value is within the normal range. The alpha1,3GT knockout- and antibiotic-re...Continue Reading

References

Oct 1, 1994·Immunological Reviews·M S Sandrin, I F McKenzie
Apr 1, 1996·Molecular and Cellular Biology·A Abuin, A Bradley
Feb 27, 1997·Nature·I WilmutK H Campbell
Dec 16, 1998·Science·Y KatoY Tsunoda
Nov 26, 1999·Biochimica Et Biophysica Acta·D H Joziasse, R Oriol
Mar 25, 2000·Theriogenology·S L SticeJ J Kane
Aug 19, 2000·Science·A OnishiA C Perry
Feb 14, 2002·Differentiation; Research in Biological Diversity·Y Tsunoda, Y Kato
Dec 21, 2002·Science·Carol J PhelpsDavid L Ayares
Jun 20, 2003·Reproduction : the Official Journal of the Society for the Study of Fertility·Chris Denning, Helen Priddle
Sep 26, 2003·Transplantation·Yutaka SendaiYoshito Aoyagi
May 5, 2004·Proceedings of the National Academy of Sciences of the United States of America·Donna Kolber-SimondsRobert J Hawley

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Citations

May 7, 2009·Zygote : the Biology of Gametes and Early Embryos·Atsushi SugawaraEimei Sato
Apr 11, 2012·Acta Histochemica Et Cytochemica·Takami TakizawaToshihiro Takizawa
Jul 17, 2009·Biotechnology Journal·Götz Laible, Lucía Alonso-González
Jan 19, 2015·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Zhongde Wang
Apr 28, 2016·Scientific Reports·Atsushi IdetaYutaka Sendai
Oct 12, 2010·Reproduction : the Official Journal of the Society for the Study of Fertility·Séverine A DegrelleIsabelle Hue

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