Selection of the Most Potent Promoter to Obtain Persistent and Prolonged Expression of the shRNAs
2:01
Preparation of Pooled Lentiviral Human Epigenetic Factor shRNA Library
4:48
Estimation of the Transduction Efficiency of Lentiviruses
6:03
Transduction of Pooled Epigenetic shRNA Library in the Drug-Resistant Cell Line
7:20
Enrichment of GFP Positive Cells
7:54
Dropout Screening to Identify Epigenetic Factors Mediating Drug Resistance
9:20
Amplification of the Integrated shRNAs by PCR
11:08
Next-Generation Sequencing and Data Analysis
11:57
Representative Results
14:30
Conclusion
TRANSCRIÇÃO
The overall goal of the following video is to demonstrate the use of the pooled shRNA epigenetic library to conduct a negative selection screening. This screening enables to identify specific epigenetic targets through sequencing. This procedure c
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High-throughput RNA interference (RNAi) screening using a pool of lentiviral shRNAs can be a tool to detect therapeutically relevant synthetic lethal targets in malignancies. We provide a pooled shRNA screening approach to investigate the epigenetic effectors in acute myeloid leukemia (AML).