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Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome
1Clinical Research Division,Fred Hutchinson Cancer Research Center, 2Hematology Branch, National Heart, Lung and Blood Institute,National Institutes of Health, 3Epigenetics Program, Department of Cell and Developmental Biology,University of Pennsylvania Perelman School of Medicine, 4Departments of Medicine and Biochemistry,University of Washington
Capitoli
- 00:04Titolo
- 01:17Selecting Single Cell Clones and Characterizing MeCP2 Reporter Expression
- 04:31Screening the Library for Reactivation Using Hygromycin B Selection
- 06:44Validating the Identified Hairpins ( Hits )
- 09:27Results: Isolation of Clones with the Reporter on the Inactive X Chromosome and Characterization of Hygromycin Resistance
- 10:17Conclusion
We report a small hairpin RNA (shRNA) and next generation sequencing-based protocol for identifying regulators of X-chromosome inactivation in a murine cell line with firefly luciferase and hygromycin resistance genes fused to the methyl CpG binding protein 2 (MeCP2) gene on the inactive X chromosome.
