Document Type
Article
Publication Date
9-23-2023
Department
Department of Biological Sciences
Abstract
MicroRNAs (miRNAs) are small non-coding RNA molecules that play a crucial role in gene regulation. They are produced through an enzyme-guided process called dicing and have an asymmetrical structure with two nucleotide overhangs at the 3' ends. Artificial microRNAs (amiRNAs or amiRs) are designed to mimic the structure of miRNAs and can be used to silence specific genes of interest. Traditionally, amiRNAs are designed based on an endogenous miRNA precursor with certain mismatches at specific positions to increase their efficiency. In this study, the authors modified the highly expressed miR168a in Arabidopsis thaliana by replacing the single miR168 stem-loop / duplex with tandem asymmetrical amiRNA duplexes that follow the statistical rules of miRNA secondary structures. These tandem amiRNA duplexes, called "two-hit" amiRNAs, were shown to have a higher efficiency in silencing GFP and endogenous PDS reporter genes compared to traditional "one-hit" amiRNAs. The authors also demonstrated the effectiveness of "two-hit" amiRNAs in silencing genes involved in miRNA, tasiRNA, and hormone signalling pathways, individually or in families. Importantly, "two-hit" amiRNAs were also able to over-express endogenous miRNAs for their functions. The authors compare "two-hit" amiRNA technology with CRISPR/Cas9 and provide a web-based amiRNA designer for easy design and wide application in plants and even animals.
Publication Title
Plant biotechnology journal
Recommended Citation
Teotia, S.,
Wang, X.,
Zhou, N.,
Wang, M.,
Liu, H.,
Qin, J.,
Han, D.,
Li, C.,
Li, C. E.,
Pan, S.,
Tang, H.,
Kang, W.,
Zhang, Z.,
Tang, X.,
Peng, T.,
&
Tang, G.
(2023).
A high-efficiency gene silencing in plants using two-hit asymmetrical artificial MicroRNAs.
Plant biotechnology journal,
21(9), 1799-1811.
http://doi.org/10.1111/pbi.14091
Retrieved from: https://digitalcommons.mtu.edu/michigantech-p/17358
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Version
Publisher's PDF
Publisher's Statement
ª2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. Publisher’s version of record: https://doi.org/10.1111/pbi.14091