Journal
/
/
Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System
DOI:

06:13 min

July 31, 2019

, , ,

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

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.

Related Videos

Read Article