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