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Generating Transgenics and Knockouts in Strongyloides Species by Microinjection
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Generating Transgenics and Knockouts in Strongyloides Species by Microinjection

Generating Transgenics and Knockouts in Strongyloides Species by Microinjection

DOI:
10.3791/63023-v

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09:42 min

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October 07, 2021

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,
1Department of Microbiology, Immunology, and Molecular Genetics,University of California, Los Angeles, 2Molecular Biology Institute,University of California, Los Angeles

Capitoli

  • 00:05Introduction
  • 00:37Preparation of Microinjection Slides
  • 01:19Collection of Young Adult Strongyloides for Microinjection
  • 03:19Preparing the Microscope and Breaking the Needle
  • 05:27Microinjecting Strongyloides
  • 07:22Collecting and Screening F1 Larvae to Recover Transgenics/Knockouts
  • 08:31Results: Generating Transgenics and Knockouts in Strongyloides Species Using Intragonadal Microinjection
  • 09:04Conclusion

Summary

Traduzione automatica

Traduzione automatica

The functional genomic toolkit for the parasitic nematodes Strongyloides stercoralis and Strongyloides ratti includes transgenesis, CRISPR/Cas9-mediated mutagenesis, and RNAi. This protocol will demonstrate how to use intragonadal microinjection to introduce transgenes and CRISPR components into S. stercoralis and S. ratti.

Tags

Keywords: MicroinjectionTransgenicsKnockoutsStrongyloidesParasitesWormsBaermannFecal-charcoalNematode Growth MediumE. Coli HB101

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