Grants and Contracts Details
Description
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.
Status | Finished |
---|---|
Effective start/end date | 1/1/22 → 5/31/24 |
Funding
- University of Florida: $160,000.00
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