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Construction and application of star polycation nanocarrier-based microRNA delivery system in Arabidopsis and maize
Jia Yang1, Shuo Yan2, Shipeng Xie1
1State Key Laboratory of Plant Physiology and Biochemistry, Engineering Research Center of Plant Growth Regulator, Ministry of Education & College of Agronomy and Biotechnology, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, People's Republic of China.
Background:
MicroRNA (miRNA) plays vital roles in the regulation of both plant architecture and stress resistance through cleavage or translation inhibition of the target messenger RNAs (mRNAs). However, miRNA-induced gene silencing remains a major challenge in vivo due to the low delivery efficiency and instability of miRNA, thus an efficient and simple method is urgently needed for miRNA transformation. Previous researches have constructed a star polycation (SPc)-mediated transdermal double-stranded RNA (dsRNA) delivery system, achieving efficient dsRNA delivery and gene silencing in insect pests.
Results:
Here, we tested SPc-based platform for direct delivery of double-stranded precursor miRNA (ds-MIRNA) into protoplasts and plants. The results showed that SPc could assemble with ds-MIRNA through electrostatic interaction to form nano-sized ds-MIRNA/SPc complex. The complex could penetrate the root cortex and be systematically transported through the vascular tissue in seedlings of Arabidopsis and maize. Meanwhile, the complex could up-regulate the expression of endocytosis-related genes in both protoplasts and plants to promote the cellular uptake. Furthermore, the SPc-delivered ds-MIRNA could efficiently increase mature miRNA amount to suppress the target gene expression, and the similar phenotypes of Arabidopsis and maize were observed compared to the transgenic plants overexpressing miRNA.
Conclusion:
To our knowledge, we report the first construction and application of star polycation nanocarrier-based platform for miRNA delivery in plants, which explores a new enable approach of plant biotechnology with efficient transformation for agricultural application.
Insights
Star polycations deliver double-stranded precursor microRNAs (ds-miRNAs) into plant cells, enabling efficient gene silencing and altered plant phenotypes. This novel method overcomes challenges in microRNA transformation for agricultural applications.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) regulate plant architecture and stress resistance by targeting messenger RNAs (mRNAs).
- Efficient in vivo miRNA delivery for gene silencing remains challenging due to low efficiency and instability.
- Previous studies utilized star polycations (SPc) for double-stranded RNA (dsRNA) delivery in insects.
Purpose of the Study:
- To develop and evaluate a star polycation (SPc)-based platform for direct delivery of double-stranded precursor microRNA (ds-miRNA) into plant protoplasts and whole plants.
- To assess the efficiency of ds-miRNA delivery, cellular uptake, and subsequent gene silencing in plants.
- To explore the potential of this platform for agricultural applications.
Main Methods:
- Formation of nano-sized ds-miRNA/SPc complexes via electrostatic interaction.
- Delivery of ds-miRNA/SPc complexes into Arabidopsis and maize protoplasts and seedlings.
- Assessment of systemic transport through vascular tissues.
- Analysis of endocytosis-related gene expression to evaluate cellular uptake.
- Quantification of mature miRNA levels and target gene suppression.
- Phenotypic analysis of treated plants compared to transgenic controls.
Main Results:
- SPc efficiently formed stable nano-complexes with ds-miRNA.
- The ds-miRNA/SPc complexes were systematically transported in Arabidopsis and maize seedlings.
- Cellular uptake was enhanced by up-regulating endocytosis-related genes.
- Delivery of ds-miRNA led to increased mature miRNA levels and suppressed target gene expression.
- Plants treated with ds-miRNA/SPc exhibited phenotypes similar to miRNA-overexpressing transgenic plants.
Conclusions:
- This study reports the first successful application of a star polycation nanocarrier platform for miRNA delivery in plants.
- The SPc-based system provides an efficient and novel approach for plant transformation and gene regulation.
- This technology holds significant potential for advancing plant biotechnology and agricultural applications.
