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- W2034881909 abstract "Self-complementary chimeric oligonucleotides (COs) composed of DNA and modified RNA residues were evaluated as a means to ( i ) create stable, site-specific base substitutions in a nuclear gene and ( ii ) introduce a frameshift in a nuclear transgene in plant cells. To demonstrate the creation of allele-specific mutations in a member of a gene family, COs were designed to target the codon for Pro-196 of SuRA, a tobacco acetolactate synthase (ALS) gene. An amino acid substitution at Pro-196 of ALS confers a herbicide-resistance phenotype that can be used as a selectable marker in plant cells. COs were designed to contain a 25-nt homology domain comprised of a five-deoxyribonucleotide region (harboring a single base mismatch to the native ALS sequence) flanked by regions each composed of 10 ribonucleotides. After recovery of herbicide-resistant tobacco cells on selective medium, DNA sequence analyses identified base conversions in the ALS gene at the codon for Pro-196. To demonstrate a site-specific insertion of a single base into a targeted gene, COs were used to restore expression of an inactive green fluorescent protein transgene that had been designed to contain a single base deletion. Recovery of fluorescent cells confirmed the deletion correction. Our results demonstrate the application of a technology to modify individual genetic loci by catalyzing either a base substitution or a base addition to specific nuclear genes; this approach should have great utility in the area of plant functional genomics." @default.
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- W2034881909 date "1999-07-20" @default.
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- W2034881909 title "A tool for functional plant genomics: Chimeric RNA/DNA oligonucleotides cause <i>in vivo</i> gene-specific mutations" @default.
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- W2034881909 doi "https://doi.org/10.1073/pnas.96.15.8774" @default.
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