Effects of Time of Deoxyribonucleic Acid Microinjection on Gene Detection and In Vitro Development of Bovine Embryos

A. E.T. Sparks, R. S. Canseco, C. G. Russell, J. L. Johnson, H. D. Moll, W. H. Velander, F. C. Gwazdauskas

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4 Scopus citations

Abstract

In vivo fertilized embryos were surgically collected from superovulated dairy cows to evaluate microinjection on embryo development and utilized the polymerase chain reaction technique for selection of transgenic embryos. Seventy-two percent of the embryos with visible pronuclei or nuclei were microinjected with DNA, and the remaining 28% served as uninjected controls. All embryos were cocultured with bovine oviductal epithelial cells. Mean final development scores of embryos within the same initial cell stage at collection were unaffected by microinjection. After 144 h of culture, 45% of the microinjected embryos developed to the morula or blastocyst stage. The transgene was detected in 50, 10, and 9% of demimorulae from embryos microinjected at the 1-, 2-, and 4-cell stages. Frequency of transgene detection was higher in morulae from 1-cell embryos than in morulae from 2- and 4-cell embryos. Use of in vitro coculture, embryo bisection, and polymerase chain reaction technique facilitated selection of bovine embryos that carried the transgene.

Original languageEnglish (US)
Pages (from-to)718-724
Number of pages7
JournalJournal of Dairy Science
Volume77
Issue number3
DOIs
Publication statusPublished - Jan 1 1994

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Keywords

  • OEC
  • PCR
  • bovine embryo culture
  • deoxyribonucleic acid microinjection
  • oviductal epithelial cells
  • polymerase chain reaction

ASJC Scopus subject areas

  • Food Science
  • Animal Science and Zoology
  • Genetics

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