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Lentiviral vektor Platform for effektiv levering af Epigenome-redigeringsværktøjer i menneskelige induceret pluripotente stamceller-afledt sygdomsmodeller
JoVE Journal
Genetics
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JoVE Journal Genetics
Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models
DOI:

13:47 min

March 29, 2019

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Chapters

  • 00:04Title
  • 02:01Plasmid Design and Construction
  • 03:20HEK-293T Cells Culturing, Plating, and Transfection
  • 05:15Virus Harvesting and Viral Particles Concentrating
  • 08:16Transduction of MD NPCs
  • 09:48Analysis of Methylation Changes
  • 11:04Results: Assessment of the LV-dCas9-DNMT3A-GFP/Puro Vectors Compared to the Naive GFP Counterpart
  • 12:45Conclusion

Summary

Automatic Translation

Målrettet DNA epigenome redigering repræsenterer en kraftfuld terapeutisk tilgang. Denne protokol beskriver produktion, oprensning og koncentration af All-in-one lentiviral vektorer husly CRISPR-dCas9-DNMT3A transgen til epigenome-redigering applikationer i menneskelige inducerede pluripotente stamceller (hiPSC)-afledt neuroner.

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