Study of Gene Silencing by Artificial miRNA

Abstract

Abstract: Artificial miRNA(amiRNA)is an effective gene silencing technology. The basic principle of amiRNA is based on its biogenesis mechanism. Using endogenous miRNA precursor as a basic framework, miRNA/miRNA* is then replaced by amiRNA/amiRNA*. The produced amiRNA is expected to silence the target gene. In this paper, the basic principle and method of amiRNA construction was introduced. AmiRNAs were designed to the target gene MYB28 as an example. Three amiRNAs respectively target to the 3' terminal region, 5' region and middle region of MYB28 coding sequence were designed. Precursor of miRNA 319a in Arabidopsis thaliana was taken as a framework and miRNA319a sequence was replaced by amiRNAs targeting to MYB28 gene. The expression level of MYB28 in transgenic plants decreased 86%. Based on our experiments, the unique characteristics of amiRNA and its further applications were discussed.

Key words: Artificial miRNA RNAi, Arabidopsis thaliana, Gene silencing

Zhang Xianhe, Kong Wenwen, Li Yong, Li Jing. Study of Gene Silencing by Artificial miRNA[J]. Biotechnology Bulletin, 2014, 0(4): 50-56.

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