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. 2021 Sep 20;12:744357. doi: 10.3389/fpls.2021.744357

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Copyright © 2021 Chen, Teotia, Lan and Tang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Techniques for the functional analysis of miRNAs in plants. (A) Transgenesis strategies to identify the functions of miRNAs via gain-of-function, overexpression of miRNAs by pre-miRNA, cDNA of miRNA or amiRNA, overexpression of miRNA cleavage-resistant target genes to mimic the effect of downregulating the expression of miRNA. (B) Techniques targeting miRNA genes: CRISPR-associated nuclease 9 (CRISPR/Cas9) system and TALEN for miRNA gene editing, CRISPR/dCas9 system for transcriptional regulation by targeting the promoter region. (C) Techniques targeting pri-/pre-miRNA: RNAi, amiRNA, and the CRISPR/Cas13a system. (D) Techniques targeting mature miRNA: AMO, TM, short tandem target mimic (STTM), and SP. AGO1, ARGONAUTE1; DCL1, DICER-LIKE1; HEN1, HUA ENHANCER1; HYL1, HYPONASTIC LEAVES1; RISC: RNA-induced silencing complex; SE, SERRATE.