Lack of RNA-DNA oligonucleotide (chimeraplast) mutagenic activity in mouse embryos

Aristides D Tagalakis, James S Owen, J Paul Simons

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

There are numerous reports of the use of RNA-DNA oligonucleotides (chimeraplasts) to correct point mutations in vitro and in vivo, including the human apolipoprotein E gene (ApoE). Despite the absence of selection for targeting, high efficiency conversion has been reported. Although mainly used to revert deleterious mutations for gene therapy applications, successful use of this approach would have the potential to greatly facilitate the production of defined mutations in mice and other species. We have attempted to create a point mutation in the mouse ApoE gene by microinjection of chimeraplast into the pronuclei of 1-cell mouse eggs. Following transfer of microinjected eggs we analysed 139 E12.5 embryos, but obtained no evidence for successful conversion.

Original languageEnglish
Pages (from-to)140-4
Number of pages5
JournalMolecular Reproduction and Development
Volume71
Issue number2
DOIs
Publication statusPublished - 24 Mar 2005

Fingerprint

Oligonucleotides
Embryonic Structures
Apolipoproteins E
RNA
Point Mutation
Eggs
DNA
Genes
Mutation
Microinjections
Genetic Therapy
In Vitro Techniques

Keywords

  • Animals
  • Apolipoproteins E/genetics
  • Cleavage Stage, Ovum/physiology
  • Female
  • Gene Targeting
  • Mutagenesis, Site-Directed
  • Oligodeoxyribonucleotides/genetics
  • Oligoribonucleotides/genetics

Cite this

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abstract = "There are numerous reports of the use of RNA-DNA oligonucleotides (chimeraplasts) to correct point mutations in vitro and in vivo, including the human apolipoprotein E gene (ApoE). Despite the absence of selection for targeting, high efficiency conversion has been reported. Although mainly used to revert deleterious mutations for gene therapy applications, successful use of this approach would have the potential to greatly facilitate the production of defined mutations in mice and other species. We have attempted to create a point mutation in the mouse ApoE gene by microinjection of chimeraplast into the pronuclei of 1-cell mouse eggs. Following transfer of microinjected eggs we analysed 139 E12.5 embryos, but obtained no evidence for successful conversion.",
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Lack of RNA-DNA oligonucleotide (chimeraplast) mutagenic activity in mouse embryos. / Tagalakis, Aristides D; Owen, James S; Simons, J Paul.

In: Molecular Reproduction and Development, Vol. 71, No. 2, 24.03.2005, p. 140-4.

Research output: Contribution to journalArticle

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AB - There are numerous reports of the use of RNA-DNA oligonucleotides (chimeraplasts) to correct point mutations in vitro and in vivo, including the human apolipoprotein E gene (ApoE). Despite the absence of selection for targeting, high efficiency conversion has been reported. Although mainly used to revert deleterious mutations for gene therapy applications, successful use of this approach would have the potential to greatly facilitate the production of defined mutations in mice and other species. We have attempted to create a point mutation in the mouse ApoE gene by microinjection of chimeraplast into the pronuclei of 1-cell mouse eggs. Following transfer of microinjected eggs we analysed 139 E12.5 embryos, but obtained no evidence for successful conversion.

KW - Animals

KW - Apolipoproteins E/genetics

KW - Cleavage Stage, Ovum/physiology

KW - Female

KW - Gene Targeting

KW - Mutagenesis, Site-Directed

KW - Oligodeoxyribonucleotides/genetics

KW - Oligoribonucleotides/genetics

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