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Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

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Updated: Jul 1, 2026

Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis

Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis

Published on: December 4, 2020

RNAi-based functional genomics tools for the beet leafhopper using microinjection and nanoparticle-based topical

Jinlong Han^1,2, Maria Paula Mejia Alonzo¹, Meihua Cui¹

¹Department of Agricultural Biology, Colorado State University, Fort Collins, Colorado, USA.

|June 30, 2026

Summary

RNA interference (RNAi) effectively targets genes in the beet leafhopper (BLH), a pest transmitting beet curly top virus (BCTV). This study establishes RNAi as a tool for BLH functional genomics and sustainable pest management.

Keywords:

Neoaliturus tenellus gene silencing gustavus microinjection nanoparticle pumilio

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Published on: May 29, 2020

Area of Science:

Agricultural Entomology
Molecular Entomology
Plant-Insect Interactions

Background:

The beet leafhopper (BLH), Neoaliturus tenellus, is a significant agricultural pest and vector for beet curly top virus (BCTV), causing substantial crop losses in the Western U.S.
Limited functional genomics tools hinder understanding of BLH survival, reproduction, and virus transmission mechanisms.

Purpose of the Study:

To evaluate the efficacy of RNA interference (RNAi) in BLH for functional genomics.
To investigate the roles of Gus and Pum3 genes in BLH fitness and BCTV interaction.
To establish microinjection and nanoparticle-mediated RNAi as viable tools for BLH research and pest management.

Main Methods:

Delivery of double-stranded RNA (dsRNA) targeting Gus and Pum3 genes via microinjection and nanoparticle-mediated topical spray.
Quantification of gene knockdown using quantitative PCR.
Assessment of BLH survival and fecundity following gene silencing.

Main Results:

Microinjection achieved >90% knockdown of Gus and Pum3 genes within 5 days.
Silencing Pum3 significantly reduced BLH survival and fecundity; silencing Gus reduced fecundity but not survival.
Nanoparticle-mediated dsRNA spray effectively reduced Gus (88%) and Pum3 (62%) expression, yielding comparable biological effects to microinjection.

Conclusions:

This study demonstrates the first successful application of microinjection and nanoparticle-mediated RNAi in BLH.
Gus and Pum3 play crucial roles in BLH fitness, impacting survival and reproduction.
These RNAi methods provide robust platforms for BLH functional genomics and developing sustainable pest control strategies.