IUCRC at the University of Florida: Improving Genome Editing in Hemipteran Pests

Grants and Contracts Details


ABSTRACT Successful application of the CRISPR/Cas9 system has been achieved in insects belonging to orders Diptera, Coleoptera, Lepidoptera, Orthoptera, and Hemiptera. Although this technology was used to edit genes in some hemipterans, target gene knockout efficiency is low. The low efficiency of gene knockouts could be due to many reasons, including the delivery of CRISPR/Cas9 to the target cell nuclei. ReMOT Control (Receptor-Mediated Ovary Transduction of Cargo) has been successfully used to deliver CRISPR/Cas9 to whitefly and other insects (1, 2). Hints at the functioning of this method in the neotropical stink bug, Euschistus heros, has also been demonstrated (3). A previous CAMTech funded project made significant progress in optimizing a gene editing method for the southern green stink bug (SGSB), Nezara viridula. The white gene was identified as a suitable marker for the identification of mutants in this insect. However, further optimization of this method for improving knockout efficiency in hemipteran insects is needed. The main objective of this project is to develop methods to improve knockout efficiency in hemipteran pests. We will use hemipteran pest species of interest to the IAB and investigate different approaches, including optimization of ReMOT Control method for delivery of CRSPR/Cas9, nanoformulation-mediated delivery and transgenic CRISPR. DNA [plasmid DNA constructs expressing single guide RNA (sgRNA) and Cas9], RNA (Cas9 mRNA/sgRNA) and Cas9/gRNA ribonucleoprotein particles (RNP) will be nanoformulated to improve the delivery of these materials and genome editing in hemipteran insects. We will also investigate the possibility of using the transgenic CRISPR in hemipterans by developing methods for the transformation of stink bugs. The delivery of piggyBac transformation system components, including enhanced transposase and reporter gene to germ cells through injection of nanoformulated material into female adults, will be evaluated.
Effective start/end date1/1/225/31/24


  • University of Florida: $160,000.00


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