Single tube, high throughput cloning of inverted repeat constructs for double-stranded RNA expression

• Published in: PloS one Vol. 4; no. 9; p. e7205
• Main Authors: Hauge, Brian, Oggero, Christopher, Nguyen, Nicole, Fu, Changlin, Dong, Fenggao
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.09.2009; Public Library of Science (PLoS)
• Subjects: Annealing; Antisense RNA; Arabidopsis - genetics; Assembly; Biotechnology; Biotechnology/Plant Biotechnology; Cloning; Cloning, Molecular; Deoxyribonucleic acid; DNA; DNA polymerase; Double-stranded RNA; Drug discovery; Electrophoresis, Agar Gel - methods; Enzymes; Escherichia coli - genetics; Fragments; Gene expression; Gene Silencing; Genetic research; Genetic Techniques; Genetic Vectors; Genetics and Genomics; Genetics and Genomics/Chromosome Biology; Genetics and Genomics/Epigenetics; Genetics and Genomics/Functional Genomics; Genetics and Genomics/Gene Discovery; Genetics and Genomics/Gene Expression; Genetics and Genomics/Gene Function; Genetics and Genomics/Genomics; Genetics and Genomics/Plant Genetics and Gene Expression; Genetics and Genomics/Plant Genomes and Evolution; Genomes; Genomics; High-throughput screening; In vivo methods and tests; Inverted repeat; Ligases; Molecular biology; Mutagenesis; Oligonucleotides, Antisense - genetics; Plants (botany); Plants, Genetically Modified - genetics; Polymerase chain reaction; Polymerase Chain Reaction - methods; Ribonucleic acid; RNA; RNA - metabolism; RNA Interference; RNA, Double-Stranded - genetics; RNA-mediated interference; Uracil - chemistry; United States > US; Missouri

RNA interference (RNAi) has emerged as a powerful tool for the targeted knockout of genes for functional genomics, system biology studies and drug discovery applications. To meet the requirements for high throughput screening in plants we have developed a new method for the rapid assembly of inverte...