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Chapters
- 00:00Introduction
- 00:24Selection of the Most Potent Promoter to Obtain Persistent and Prolonged Expression of the shRNAs
- 02:01Preparation of Pooled Lentiviral Human Epigenetic Factor shRNA Library
- 04:48Estimation of the Transduction Efficiency of Lentiviruses
- 06:03Transduction of Pooled Epigenetic shRNA Library in the Drug-Resistant Cell Line
- 07:20Enrichment of GFP Positive Cells
- 07:54Dropout Screening to Identify Epigenetic Factors Mediating Drug Resistance
- 09:20Amplification of the Integrated shRNAs by PCR
- 11:08Next-Generation Sequencing and Data Analysis
- 11:57Representative Results
- 14:30Conclusion
High-throughput RNA-interferentie (RNAi) screening met behulp van een pool van lentivirale shRNA's kan een hulpmiddel zijn om therapeutisch relevante synthetische dodelijke doelwitten bij maligniteiten te detecteren. We bieden een gepoolde shRNA-screeningsbenadering om de epigenetische effectoren bij acute myeloïde leukemie (AML) te onderzoeken.
