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Using CRISPR/Cas9 to identify genes required for mechanosensory neuron development and function.

Christopher J Johnson1, Akhil Kulkarni1, William J Buxton1

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Summary

Researchers screened tunicates for genes involved in metamorphosis using CRISPR technology. They identified Vamp1/2/3 as crucial for sensory neurons in the tunicate larva to trigger settlement and metamorphosis.

Keywords:
AscidianCRISPR/Cas9CionaEducationMarine larvaeTunicate

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Area of Science:

  • Marine biology
  • Developmental biology
  • Neuroscience

Background:

  • Tunicates are marine invertebrates and the closest relatives to vertebrates.
  • Tunicate larvae exhibit a biphasic life cycle with a swimming larva and a sessile adult.
  • Understanding tunicate metamorphosis is key to understanding vertebrate evolution.

Purpose of the Study:

  • To screen for genes involved in mechanosensation and metamorphosis in tunicate larvae.
  • To utilize CRISPR/Cas9 technology in an undergraduate research setting.
  • To identify novel genes regulating the settlement and metamorphosis process in tunicates.

Main Methods:

  • Tissue-specific CRISPR/Cas9-mediated mutagenesis was employed in Ciona robusta.
  • A genetic screen was conducted within an undergraduate capstone research course.
  • The study focused on sensory neurons in the anterior papillae of tunicate larvae.

Main Results:

  • A screen identified at least one gene, Vamp1/2/3, essential for papillae-mediated metamorphosis.
  • The study provides protocols for replicating CRISPR-based experiments in educational settings.
  • Mechanosensory transduction in tunicate papillae is crucial for initiating metamorphosis.

Conclusions:

  • Vamp1/2/3 plays a critical role in the tunicate larval metamorphosis process.
  • CRISPR technology can be effectively integrated into undergraduate research for gene discovery.
  • This work facilitates further investigation into the neural basis of tunicate settlement and metamorphosis.