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Triagem de biblioteca shRNA agrupada para identificar fatores que modulam um fenótipo de resistência a drogas
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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
A triagem de interferência de RNA de alto rendimento (RNAi) usando um pool de shRNAs lentiviral pode ser uma ferramenta para detectar alvos letais sintéticos terapeuticamente relevantes em malignidades. Fornecemos uma abordagem de triagem de shRNA agrupada para investigar os efeitos epigenéticos na leucemia mielóide aguda (LMA).
