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Updated: Jan 21, 2026

Culturing and Genetically Manipulating Entomopathogenic Nematodes
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Culturing and Genetically Manipulating Entomopathogenic Nematodes

Published on: March 31, 2022

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CRISPR-Cas9-based Mutagenesis in the Entomopathogenic Nematode Steinernema hermaphroditum and the Maintenance of

Sally W Ireri1, Mengyi Cao2

  • 1Division of Biosphere Sciences and Engineering, Carnegie Institution for Science.

Journal of Visualized Experiments : Jove
|January 19, 2026
PubMed

Insights

Scientists developed a CRISPR-Cas9 genome editing tool for entomopathogenic nematodes (EPNs). This new genetic tool in Steinernema hermaphroditum enables targeted gene modification for studying nematode-bacterium interactions and agricultural applications.

Area of Science:

  • Molecular Biology
  • Genetics
  • Nematology

Background:

  • Entomopathogenic nematodes (EPNs) form symbiotic relationships with bacteria like Xenorhabdus and Photorhabdus.
  • These nematode-bacterium pairs are crucial for controlling insect pests, particularly Lepidoptera and Coleoptera larvae.
  • Developing stable genetic tools for EPNs is essential for understanding their mutualistic and parasitic interactions.

Purpose of the Study:

  • To establish a reliable CRISPR-Cas9 genome editing platform in the entomopathogenic nematode Steinernema hermaphroditum.
  • To enable stable and transgenerational genetic manipulation in EPNs for research purposes.
  • To facilitate the study of molecular mechanisms underlying EPN-bacterium mutualism and parasitism.

Main Methods:

  • Utilized CRISPR-Cas9 ribonucleoprotein complexes delivered via gonadal microinjection in Steinernema hermaphroditum.
  • Developed a protocol for efficient, targeted gene disruption in EPNs.
  • Leveraged the hermaphroditic reproduction of S. hermaphroditum to streamline mutant line generation.

Main Results:

  • Successfully demonstrated CRISPR-Cas9 genome editing in Steinernema hermaphroditum.
  • Generated a characteristic twitching phenotype by modifying the unc-22 gene, validating targeted mutagenesis.
  • Established a functional genetic platform for S. hermaphroditum, including potential for transgene expression.

Conclusions:

  • The developed CRISPR-Cas9 platform provides a robust method for genetic manipulation in Steinernema hermaphroditum.
  • This breakthrough enables future research into EPN biology, mutualism, and parasitism.
  • The framework can be extended to other agriculturally and ecologically important EPN species.

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