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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

23
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Related Experiment Video

Updated: Jul 12, 2025

Digital-Droplet PCR to Detect Indels Mutations in Genetically Modified Anopheline Mosquito Populations
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CRISPR-based diagnostics detects invasive insect pests.

Pathour R Shashank1,2, Brandon M Parker1,3,4, Santosh R Rananaware5

  • 1McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, Florida, USA.

Molecular Ecology Resources
|October 27, 2023
PubMed
Summary
This summary is machine-generated.

A new CRISPR-based diagnostic tool accurately identifies invasive moth species using the CO1 gene. This rapid, sensitive method offers a revolutionary approach to organism detection and monitoring in various biological and medical fields.

Keywords:
Keiferia lycopersicellaPhthorimaea absolutaScrobipalpa atriplicellaTuta absolutaCRISPRCas12aRPAdiagnosticsgenetic identificationleaf miner

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Area of Science:

  • Molecular Biology
  • Genetics
  • Ecology

Background:

  • Rapid organism identification is crucial for ecological studies, disease diagnosis, and pest management.
  • CRISPR-based diagnostics present a promising, accurate, and fast alternative to traditional identification methods.

Purpose of the Study:

  • To develop and validate a CRISPR-based diagnostic assay for rapid and accurate identification of animal species.
  • To utilize the universal cytochrome-oxidase 1 (CO1) gene for broad applicability across Animalia.
  • To test the assay's efficacy on three globally significant invasive moth pests.

Main Methods:

  • Developed a CRISPR-based diagnostic assay targeting the cytochrome-oxidase 1 (CO1) gene.
  • Combined recombinase polymerase amplification (RPA) with CRISPR for enhanced signal generation.
  • Tested the assay on three invasive moth species: Keiferia lycopersicella, Phthorimaea absoluta, and Scrobipalpa atriplicella.

Main Results:

  • Achieved 100% accuracy in identifying all three tested moth species.
  • Demonstrated high sensitivity with detection limits as low as 120 fM for P. absoluta.
  • The assay is significantly more sensitive than real-time PCR and requires minimal laboratory equipment.

Conclusions:

  • The developed CRISPR-RPA assay provides a rapid, sensitive, and accurate method for animal detection.
  • This approach has the potential to revolutionize organism identification and monitoring, especially for invasive species.
  • The method is user-friendly, reduces contamination risk, and can be completed within an hour.