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En effektiv In Vitro gennemførelse metode af en Transcriptionally regulerede sovende skønhed System pakket ind i en Integration defekte Lentiviral vektor
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JoVE Journal Biology
An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector

En effektiv In Vitro gennemførelse metode af en Transcriptionally regulerede sovende skønhed System pakket ind i en Integration defekte Lentiviral vektor

DOI:
10.3791/56742-v

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10:13 min

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January 12, 2018

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, ,
1Centre for Regenerative Medicine, Department of Life Sciences,University of Modena and Reggio Emilia

Chapters

  • 00:05Title
  • 00:51Transduction of Human Primary Keratinocytes
  • 04:31Cytofluorimetric Analysis of Transduced Primary Keratinocytes
  • 06:24Quantitative RT-PCR (qRT-PCR)
  • 07:24Results: Sleeping Beauty System Achieves Stable Integration of a GOI in the Human Genome
  • 08:40Conclusion

Summary

Automatic Translation

Automatic Translation

Denne protokol beskriver en metode til at opnå stabile integration af et gen af interesse i det menneskelige genom transcriptionally regulerede Tornerose systemet. Forberedelse af integrationen af defekte lentiviral vektorer, in vitro- transduktion af menneskelige celler og molekylær analysen på transduced celler er rapporteret.

Tags

Sleeping Beauty SystemIntegration Defective Lentiviral VectorGene TherapyPrimary CellsTranscriptionally RegulatedTransposition MethodIn VitroHyperactive SB100X TransposaseGene Of InterestTetracycline-regulatedPolybrene

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