Functional Characterization and Putative Regulatory Mechanism of an RNAi Efficiency-Related Nuclease (REase) in the...

Area of Science:

• Molecular Biology
• Insect Biochemistry
• RNA Interference

Background:

• Lepidopteran-specific RNAi efficiency-related nuclease (REase) degrades double-strand RNA (dsRNA).
• The regulatory mechanisms of REase in insects remain largely uncharacterized.

Purpose of the Study:

• To identify and characterize SfREase in Spodoptera frugiperda.
• To elucidate the regulatory mechanism and functional role of SfREase in RNAi efficiency.

Main Methods:

• Identification and characterization of SfREase in Spodoptera frugiperda.
• Analysis of SfREase expression under different conditions (dsEGFP, high temperature, starvation).
• In vitro degradation assays using recombinant SfREase and analysis of dsRNA degradation in larval fluids.
• RNA interference (RNAi) and dual-luciferase reporter assays to determine SfREase regulation by FOXO.

Main Results:

• SfREase expression is upregulated by dsEGFP and high temperature, and downregulated by starvation.
• Knockdown of SfREase significantly reduces dsRNA degradation and enhances RNAi efficiency.
• Recombinant SfREase exhibits concentration-dependent dsRNA degradation activity.
• FOXO was identified as a negative regulator of SfREase activity.

Conclusions:

• SfREase plays a crucial role in dsRNA degradation in Spodoptera frugiperda, impacting RNAi efficiency.
• The study reveals a novel regulatory mechanism involving FOXO, providing insights into REase function.
• Findings have significant implications for developing targeted RNAi-based pest control strategies for Spodoptera frugiperda.