Date of Award

2017

Document Type

Open Access Dissertation

Degree Name

Doctor of Philosophy in Biological Sciences (PhD)

Administrative Home Department

Department of Biological Sciences

Advisor 1

Guiliang Tang

Committee Member 1

Xiaoqing Tang

Committee Member 2

Haiying Liu

Committee Member 3

Michael Gretz

Abstract

MicroRNA (miRNAs) and other endogenous small RNAs play important role in regulating gene expression in eukaryotes. Till now, several techniques have been developed for miRNA function analysis. Because of the characteristic of miRNAs, the most applicable method for miRNA function research is blocking the whole miRNA family specifically. Here, we report an artificial non-coding transcript termed Short Tandem Target Mimic (STTM) for blocking miRNA activity in Arabidopsis thaliana. The STTM composed of two miRNA target sites with three nucleotides bulge in the miRNA cleavage site, the two-miRNA targets separated by spacer linker. We used this technique blocking several important endogenous miRNA with high efficiency; we also demonstrated that the degradation of miRNA induced by small RNA degradation nucleases (SDNs). In summary, we generated an effective approach to block endogenous miRNA, therefore providing a powerful tool for miRNA function research in plants.

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