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Published on: April 10, 2018
Lack of RNA-DNA oligonucleotide (chimeraplast) mutagenic activity in mouse embryos
Aristides D Tagalakis1, James S Owen, J Paul Simons
1Department of Anatomy and Developmental Biology, Hampstead Campus, University College London, London, United Kingdom.
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.
Insights
RNA-DNA oligonucleotides (chimeraplasts) failed to correct point mutations in the mouse apolipoprotein E (ApoE) gene. This study found no evidence of successful gene conversion in mouse embryos after microinjection of chimeraplasts.
Area of Science:
- Molecular Biology
- Genetics
- Gene Therapy
Background:
- RNA-DNA oligonucleotides, termed chimeraplasts, are reported to efficiently correct point mutations in vitro and in vivo.
- This technology has potential applications in gene therapy and creating defined mutations in animal models.
Purpose of the Study:
- To investigate the efficacy of chimeraplast-mediated gene correction for introducing a specific point mutation into the mouse apolipoprotein E (ApoE) gene.
- To assess the feasibility of using this method for generating genetically modified mouse models.
Main Methods:
- Microinjection of chimeraplasts into the pronuclei of 1-cell mouse embryos.
- Analysis of E12.5 mouse embryos for evidence of successful gene conversion at the targeted ApoE locus.
Main Results:
- No successful gene conversion events were detected in 139 analyzed mouse embryos.
- The microinjection of chimeraplasts did not result in the intended point mutation in the mouse ApoE gene.
Conclusions:
- The study failed to demonstrate successful gene correction using chimeraplasts in mouse embryos.
- Further optimization or alternative strategies may be required for efficient gene editing in mouse models using this approach.
