Transfer of a mutant dihydrofolate reductase gene into pre- and postimplantation mouse embryos by a replication-competent retrovirus vector. Academic Article uri icon

Overview

MeSH

  • Animals
  • Blotting, Southern
  • Cell Line
  • Female
  • Male
  • Methotrexate
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Nucleic Acid Hybridization
  • Repetitive Sequences, Nucleic Acid

MeSH Major

  • Blastocyst
  • Embryo, Mammalian
  • Genes
  • Genetic Vectors
  • Moloney murine leukemia virus
  • Mutation
  • Tetrahydrofolate Dehydrogenase
  • Transfection

abstract

  • In order to explore the potential of retrovirus vectors for efficiently transferring foreign genes into mouse embryos, a replication-competent recombinant Moloney murine leukemia virus (Mo-MLV) vector carrying a mutant dihydrofolate reductase (DHFR) cDNA insert in the U3 region of the viral long terminal repeat was used to infect pre- and postimplantation embryos. When preimplantation mouse embryos were infected with the vector, as expected, the provirus integrated into the embryos and the germ line with the same efficiency as that observed with wild-type Mo-MLV, leading to inactivation of the recombinant virus. In contrast, when postimplantation mouse embryos were microinjected with virus-producing cells, between 90 to 100% of the surviving animals proved to be infected with the virus. The recombinant virus spread as efficiently as wild-type Mo-MLV in the infected embryos, resulting in up to three to five proviral copies per genome in heart, thymus, and brain tissues. Substantial expression of mutant DHFR*-coding viral message was found in all somatic tissues analyzed, the amounts correlating with the proviral copy number in the respective organ. These results suggest that replication-competent vectors are useful for efficient transfer and expression of foreign genes into tissues or whole animals when virus spread is needed.

publication date

  • November 1989

has subject area

  • Animals
  • Blastocyst
  • Blotting, Southern
  • Cell Line
  • Embryo, Mammalian
  • Female
  • Genes
  • Genetic Vectors
  • Male
  • Methotrexate
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Moloney murine leukemia virus
  • Mutation
  • Nucleic Acid Hybridization
  • Repetitive Sequences, Nucleic Acid
  • Tetrahydrofolate Dehydrogenase
  • Transfection

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC251124

PubMed ID

  • 2795720

Additional Document Info

start page

  • 4857

end page

  • 4865

volume

  • 63

number

  • 11