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Repeat mediated excision of gene drive elements for restoring wild-type populations
Pratima R Chennuri1, Josef Zapletal2, Raquel D Monfardini1
1Department of Entomology and AgriLife Research, Texas A&M University, College Station, Texas, United States of America.
Plos Genetics
|November 7, 2024
Summary
Repeat Mediated Excision of a Drive Element (ReMEDE) uses single strand annealing for precise gene drive removal. This technology shows potential for restoring wild-type populations without environmental modification.
Area of Science:
- Genetics
- Molecular Biology
- Gene Drive Technology
Background:
- Gene drive systems can rapidly alter wild populations.
- Developing safe and reversible gene drive systems is crucial for ecological applications.
- Mitigation strategies are needed to control or reverse unintended gene drive effects.
Purpose of the Study:
- To develop and evaluate Repeat Mediated Excision of a Drive Element (ReMEDE) as a gene drive mitigation strategy.
- To demonstrate the precise autocatalytic excision of a drive element using single strand annealing (SSA).
- To assess the efficacy of ReMEDE in replacing drive alleles with wild-type alleles in Drosophila melanogaster.
Main Methods:
- Engineering direct repeats flanking a drive allele.
- Inducing a double-strand DNA break (DSB) within the drive allele.
- Incorporating ReMEDE into a mutagenic chain reaction (MCR) gene drive targeting the yellow gene.
- Analyzing gene conversion and inheritance patterns through sequencing.
Main Results:
- ReMEDE successfully replaced drive alleles with wild-type alleles in Drosophila melanogaster.
- Sequencing confirmed transgene excision by SSA, with ~4% inheritance of a silent marker.
- Observed were phenotypically wild-type flies with retained or mutated sequences, and ~14% intact paternally inherited alleles.
Conclusions:
- ReMEDE demonstrates potential as a gene drive mitigation strategy by enabling precise autocatalytic excision of drive elements.
- The technology can restore wild-type populations without additional transgenic releases.
- Further research is needed to optimize ReMEDE for broader application as a gene drive reversal tool.
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