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Cited by 27CRISPR and Genetic EngineeringPlant Genetic and Mutation StudiesTransgenic Plants and Applications

Conditional knockdown of OsMLH1 to improve plant prime editing systems without disturbing fertility in rice

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Liu, Xiaoshuang, Gu, Dongfang, Zhang, Yiru, Jiang, Yingli, Xiao, Zhi, Xu, Rongfang, Qin, Ruiying, Li, Juan +1 more·2024·Genome Biology
DOI 10.1186/s13059-024-03282-yPMC PMC11110357
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Abstract

Background: High-efficiency prime editing (PE) is desirable for precise genome manipulation. The activity of mammalian PE systems can be largely improved by inhibiting DNA mismatch repair by coexpressing a dominant-negative variant of MLH1. However, this strategy has not been widely used for PE optimization in plants, possibly because of its less conspicuous effects and inconsistent performance at different sites. Results: We show that direct RNAi knockdown of OsMLH1 in an ePE5c system increases the efficiency of our most recently updated PE tool by 1.30- to 2.11-fold in stably transformed rice cells, resulting in as many as 85.42% homozygous mutants in the T 0 generation. The high specificity of ePE5c is revealed by whole-genome sequencing. To overcome the partial sterility induced by OsMLH1 knockdown of ePE5c, a conditional excision system is introduced to remove the RNAi module by Cre-mediated site-specific recombination. Using a simple approach of enriching excision events, we generate 100% RNAi module-free plants in the T 0 generation. The increase in efficiency due to OsMLH1 knockdown is maintained in the excised plants, whose fertility is not impaired. Conclusions: This study provides a safe and reliable plant PE optimization strategy for improving editing efficiency without disturbing plant development via transient MMR inhibition with an excisable RNAi module of MLH1.

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  • The ZmGA20ox3 gene demonstrated as prominent negative regulator for enhancing drought tolerance in maize seedling, reduces Anthesis-Silking Interval (ASI) delay and decreasing the yield loss significantly in the field under drought conditions.

    “The ZmGA20ox3 gene demonstrated as prominent negative regulator for enhancing drought tolerance in maize seedling, reduces Anthesis-Silking Interval (ASI) delay and decreasing the yield loss significantly in the field under drought conditions (Liu Y. et al., 2024).”

    From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Liu Y. et al., 2024

    Split into 2 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.

    Experimental — accuracy not yet measured

  • Editing the ZmGA20ox3 gene in maize improved plant architecture and enhanced drought tolerance.

    “Liu Y. et al. (2024) demonstrated that editing the ZmGA20ox3 gene in maize improved plant architecture and enhanced drought tolerance.”

    From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Liu Y. et al., 2024

    The Results section reports editing targets in rice, not ZmGA20ox3 in maize. A search for ZmGA20ox3, maize architecture, and drought tolerance found no passage reporting the claimed effects; the paper’s drought treatment concerns excision of an editing cassette in rice. (LLM, conf=0.76)

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Cited by

Sorted by citations

Authors
Liu, X., Gu, D., Zhang, Y., Jiang, Y., Xiao, Z., Xu, R., Qin, R., Li, J., & Wei, P.
Affiliations
  • Anhui Agricultural University (CN)
  • Anhui Academy of Agricultural Sciences (CN)
Corresponding author
  • Li, Juan
  • Wei, Pengcheng
ORCID
Venue
Genome Biology
Publisher
Springer Science and Business Media LLC
Published
2024-05-21
Volume
25
Issue
1
Pages
131-131
ISSN
  • 1465-6906 (ISSN-L)
  • 1474-760X
DOI
10.1186/s13059-024-03282-y
PMC
PMC11110357
OpenAlex
W4398156486
Cited by
27
References
1
Citations by year
2026: 8 · 2025: 11 · 2024: 8
FWCI
3.68
Type
journal article
Language
en
Open access
Licence
Locations
  • Genome biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
STI-2030 (2023ZD04074)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Genetic and Mutation Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
Keywords
prime editing, rice, plant fertility, genome editing efficiency
MeSH terms
MutL Protein Homolog 1, Gene Editing, Fertility, Plant Proteins, Oryza, Plants, Genetically Modified, RNA Interference, Gene Knockdown Techniques, CRISPR-Cas Systems

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