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

By AD Tagalakis, JS Owen and JP Simons

Abstract

There are numerous reports of the use of RNA-DNA oligonucleoticles (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. (c) 2005 Wiley-Liss, Inc

Topics: gene targeting, pronuclear injection, chimeraplast, oligonucleotide, mutagenesis, SNP, TARGETED NUCLEOTIDE EXCHANGE, SINGLE-STRANDED OLIGONUCLEOTIDES, GENE REPAIR, STEM-CELLS, MAMMALIAN-CELLS, IN-VIVO, RNA/DNA OLIGONUCLEOTIDES, MUTANT MICE, GENOME-WIDE, MUTATIONS
Publisher: WILEY-LISS
Year: 2005
DOI identifier: 10.1002/mrd.20250
OAI identifier: oai:eprints.ucl.ac.uk.OAI2:303
Provided by: UCL Discovery

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