Fate of microinjected genes in preimplantation mouse embryos

Tom Burdon, R. J Wall

Research output: Contribution to journalArticlepeer-review

Abstract

The state of genes microinjected into mouse embryos was followed from the one-cell to the blastocyst stage using the polymerase chain reaction (PCR). Microinjected DNA was detected in all one-, two-, and four-cell injected embryos and in 44% of morula and 26% of blastocysts. Head-to-tail ligation of microinjected genes, a common feature of stably integrated transgene arrays, was detected in all embryos after injection of microinjected genes and occurred irrespective of the structure at the ends of the injected genes. Sensitivity of microinjected DNA to a methylation-dependent restriction endonuclease Dpn I was lost in all embryos by the two-cell stage (24 hr), indicating a change in DNA methylation, independent of transgene integration. Dissociation of blastomeres prior to compaction revealed a mosaic distribution of the microinjected DNA within the embryo and supports the notion that injected genes form a limited number of arrays, which segregate independently until they integrate into the genome or are degraded.
Original languageEnglish
Pages (from-to)436-42
Number of pages7
JournalMolecular Reproduction and Development
Volume33
Issue number4
DOIs
Publication statusPublished - Dec 1992

Keywords

  • Animals
  • Base Sequence
  • Blastocyst/metabolism
  • DNA/administration & dosage
  • DNA/genetics
  • DNA/metabolism
  • Female
  • Genetic Engineering
  • Genetic Techniques
  • Methylation
  • Mice
  • Mice, Transgenic/genetics
  • Microinjections
  • Molecular Sequence Data
  • Mosaicism
  • Polymerase Chain Reaction
  • Pregnancy
  • Transfection

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