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Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System
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Genetics
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JoVE Journal Genetics
Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

DOI:
10.3791/59962-v

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06:13 min

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July 31, 2019

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, , ,
1Biomedical Research Institute,National Institute of Advanced Industrial Science and Technology (AIST)

Chapters

  • 00:04Title
  • 00:35Differentiation of Trypomastigotes into Extracellular Amastigotes
  • 01:54Electroporation of Extracellular Amastigotes
  • 03:01Monitoring the Growth of Knockout Cells
  • 04:35Results: Growth Monitoring and Phenotypic Differences of Knockout Parasites
  • 05:41Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a protocol to introduce a gene knockout into the extracellular amastigote of Trypanosoma cruzi, using the CRISPR/Cas9 system. The growth phenotype can be followed up either by cell counting of axenic amastigote culture or by proliferation of intracellular amastigotes after host cell invasion.

Tags

Gene KnockoutCRISPR/Cas9Trypanosoma CruziAmastigoteHost Cell InfectionParasitologyExtracellular AmastigotesTrypomastigotesCell CultureElectroporation

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