Chimeric RNA/DNA oligonucleotide-directed gene targeting in rice.
Site-specific mutagenesis in a rice genome was obtained by introducing chimeric RNA/DNA oligonucleotides (COs) by means of particle bombardment. Three COs were designed to target the independent codons for Pro-171, Trp-548 and Ser-627 of the endogenous rice acetolactate synthase (ALS) gene so it would confer resistance to ALS-inhibiting herbicides. Sequencing of the ALS gene of herbicide-resistant plants demonstrated that the ALS sequence was modified in a site-specific fashion. The efficiency of gene conversion mediated by COs was estimated to be 1x10(-4). These results demonstrate that CO-directed gene targeting is feasible in rice.
Okuzaki
A
A
Laboratory of Plant Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, 981-8555 Sendai, Japan.
Toriyama
K
K
D000095:Acetolactate Synthase / Q000235:genetics; D001483:Base Sequence; D019470:Biolistics; D002678:Chimera / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D005785:Gene Conversion; D018390:Gene Targeting / Q000379:methods*; D018745:Genome, Plant; D006540:Herbicides / Q000494:pharmacology; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides / Q000235:genetics; D012275:Oryza / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D030821:Plants, Genetically Modified; D012313:RNA / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics
D018744:DNA, Plant; D004274:DNA, Recombinant; D006540:Herbicides; D009841:Oligonucleotides; D018749:RNA, Plant; C039919:RNA, recombinant; D012313:RNA; D000095:Acetolactate Synthase