# CRISPR/Cas9-Mediated Knockout of the White Gene in Agasicles hygrophila

## TL;DR

Researchers used CRISPR/Cas9 to edit the white gene in a beetle species, creating a white-eyed mutant that could help improve biological control of invasive weeds.

## Contribution

The study introduces a CRISPR/Cas9-based gene editing method for functional genomics in Agasicles hygrophila.

## Key findings

- CRISPR/Cas9 successfully induced mutations in the white gene of A. hygrophila, resulting in white eyes.
- The method provides a new genetic tool for studying and improving this biocontrol agent.
- The white-eyed mutant strain may support future transgenic research on the species.

## Abstract

Agasicles hygrophila is the most effective natural enemy for the control of the invasive weed Alternanthera philoxeroides (Mart.) Griseb. However, research on the gene function and potential genetic improvement of A. hygrophila is limited due to a lack of effective genetic tools. In this study, we employed the A. hygrophila white (AhW) gene as a target gene to develop a CRISPR/Cas9-based gene editing method applicable to A. hygrophila. We showed that injection of Cas9/sgRNA ribonucleoprotein complexes (RNPs) of the AhW gene into pre-blastoderm eggs induced genetic insertion and deletion mutations, leading to white eyes. Our results demonstrate that CRISPR/Cas9-mediated gene editing is possible in A. hygrophila, offering a valuable tool for studies of functional genomics and genetic improvement of A. hygrophila, which could potentially lead to more effective control of invasive weeds through the development of improved strains of this biocontrol agent. In addition, the white-eyed mutant strain we developed could potentially be useful for other transgenic research studies on this species.