An efficient and multiple target transgenic RNAi technique with low toxicity in Drosophila

Huan-Huan Qiao, Fang Wang, Rong-Gang Xu, Jin Sun, Ruibao Zhu, Decai Mao, Xingjie Ren, Xia Wang, Yu Jia, Ping Peng, Da Shen, Lu-Ping Liu, Zhijie Chang, Guirong Wang, Shao Li, Jun-Yuan Ji, Qingfei Liu and Jian-Quan Ni ()
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Huan-Huan Qiao: Tsinghua University
Fang Wang: Tsinghua University
Rong-Gang Xu: Tsinghua University
Jin Sun: Tsinghua University
Ruibao Zhu: Tsinghua University
Decai Mao: Tsinghua University
Xingjie Ren: University of California San Francisco
Xia Wang: Tsinghua University
Yu Jia: Tsinghua University
Ping Peng: Tsinghua University
Da Shen: Tsinghua University
Lu-Ping Liu: Tsinghua University
Zhijie Chang: Tsinghua University
Guirong Wang: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection
Shao Li: Tsinghua University
Jun-Yuan Ji: Texas A&M Health Science Center
Qingfei Liu: Tsinghua University
Jian-Quan Ni: Tsinghua University

Nature Communications, 2018, vol. 9, issue 1, 1-13

Abstract: Abstract Being relatively simple and practical, Drosophila transgenic RNAi is the technique of top priority choice to quickly study genes with pleiotropic functions. However, drawbacks have emerged over time, such as high level of false positive and negative results. To overcome these shortcomings and increase efficiency, specificity and versatility, we develop a next generation transgenic RNAi system. With this system, the leaky expression of the basal promoter is significantly reduced, as well as the heterozygous ratio of transgenic RNAi flies. In addition, it has been first achieved to precisely and efficiently modulate highly expressed genes. Furthermore, we increase versatility which can simultaneously knock down multiple genes in one step. A case illustration is provided of how this system can be used to study the synthetic developmental effect of histone acetyltransferases. Finally, we have generated a collection of transgenic RNAi lines for those genes that are highly homologous to human disease genes.

Date: 2018
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DOI: 10.1038/s41467-018-06537-y

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