Simultaneous knockdown of six non-family genes using a single synthetic RNAi fragment in Arabidopsis thaliana [electronic resource].
- Washington, D.C. : United States. Dept. of Energy. Office of Science, 2016. and Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
- Physical Description:
- Article numbers 16 : digital, PDF file
- Additional Creators:
- Oak Ridge National Laboratory, United States. Department of Energy. Office of Science, and United States. Department of Energy. Office of Scientific and Technical Information
- Restrictions on Access:
- Free-to-read Unrestricted online access
- Genetic engineering of plants that results in successful establishment of new biochemical or regulatory pathways requires stable introduction of one or more genes into the plant genome. It might also be necessary to down-regulate or turn off expression of endogenous genes in order to reduce activity of competing pathways. An established way to knockdown gene expression in plants is expressing a hairpin-RNAi construct, eventually leading to degradation of a specifically targeted mRNA. Knockdown of multiple genes that do not share homologous sequences is still challenging and involves either sophisticated cloning strategies to create vectors with different serial expression constructs or multiple transformation events that is often restricted by a lack of available transformation markers. Synthetic RNAi fragments were assembled in yeast carrying homologous sequences to six or seven non-family genes and introduced into pAGRIKOLA. Transformation of Arabidopsis thaliana and subsequent expression analysis of targeted genes proved efficient knockdown of all target genes. In conclusion, we present a simple and cost-effective method to create constructs to simultaneously knockdown multiple non-family genes or genes that do not share sequence homology. The presented method can be applied in plant and animal synthetic biology as well as traditional plant and animal genetic engineering.
- Published through SciTech Connect., 02/17/2016., ": 116", Plant Methods 12 1 ISSN 1746-4811 AM, and Olaf Czarnecki; Anthony C. Bryan; Sara S. Jawdy; Xiaohan Yang; Zong-Ming Cheng; Jin-Gui Chen; Gerald A. Tuskan.
- Funding Information:
- SC0008834 and AC05-00OR22725
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