Functional allelic diversity of PTGS components revealed by a high-throughput RUBY-based screening.

Authors

Xin Wei, Yanming Hou, Yuyang Si, Weiyi Guo, Yuping Qiu, Haofeng Li, Yunhui Sun, Zhonghua Tang, Liqiang Mu, Wei Yan, Hongwei Guo

Published in

The Plant journal : for cell and molecular biology. Volume 127. Issue 2. Pages e71041.

Abstract

Post-transcriptional gene silencing (PTGS) mediated by small RNA significantly impairs both transgene efficiency and endogenous gene expression. Although multiple PTGS components have been identified through forward genetic screening using various reporter systems, mutations reported for these known PTGS components remain limited, hindering our understanding of the underlying mechanisms and the functional roles of specific protein domains. Here, we combined the 5'-3' RNA decay mutant ein5-1 with a non-invasive RUBY system to perform high-throughput screening for mutants that can restore RUBY expression. We identified 239 mutations covering almost all critical protein domains in key PTGS components and promising novel factors. Through systematic phenotypic evaluation, we found spatial-temporal repression of PTGS in distinct mutants. Mutations affecting enzymatic/catalytic activity center, RNA-binding sites, or interactions between essential PTGS components dramatically reduced RUBY-derived siRNA production. These mutant alleles provide valuable genetic resources for investigating domain-specific functions of known PTGS factors, thereby offering support for fine-tuning PTGS efficiency.

PMID:
42460500
Bibliographic data and abstract were imported from PubMed on 16 Jul 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.