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JoVE Journal Genetics
Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome
 

Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome

Article DOI: 10.3791/56398-v • 11:15 min • March 2nd, 2018
March 2nd, 2018 •
1,2, 1, 1, 1, 1, 1, 1, 3, 1,4
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

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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.

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ShRNA Screen MeCP2 Reactivation Inactive X Chromosome Mouse Tendon Fibroblasts Cell Harvest Trypsin EDTA DMEM-10 Cell Dilution 96 Well Plates Cell Confluence Media Replacement Cell Mobilization Luciferase Assay Clone Selection Positive Control
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