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Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains
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Biologia
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JoVE Journal Biologia
Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains

Direct Protein Delivery to Mammalian Cells Using Cell-permeable Cys2-His2 Zinc-finger Domains

DOI:
10.3791/52814-v

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11:24 min

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March 25, 2015

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,
1Departments of Chemistry and Cell and Molecular Biology,The Scripps Research Institute, 2Shanghai Institute for Advanced Immunochemical Studies,ShanghaiTech University

Capítulos

  • 00:05Título
  • 01:23Cloning Emerald GFP into an Expression Vector Containing Two-finger ZiF Domains
  • 04:22Purification of the ZiF-EmGFP Fusion Protein
  • 07:17Protein Transduction of HeLa Cells
  • 10:07Results: Fusion of 2F-ZiF Domains to EmGFP Enables Delivery of EmGFP to HeLa Cells
  • 10:57Conclusion

Summary

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Zinc-finger domains are intrinsically cell-permeable and capable of mediating protein delivery into a broad range of mammalian cell types. Here, a detailed step-by-step protocol for implementing zinc-finger technology for intracellular protein delivery is presented.

Tags

Zinc-fingerProtein DeliveryCell-permeableCys2-His2Genome EngineeringIntracellular DeliveryCell MembraneProtein TransductionEnzyme ActivityCytosolic DeliveryMammalian Cells

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