Profiling of gene expression or chromatin marks in specific cell populations usually requires that the cells be isolated physically. Southall et al. circumvented this need by modifying the DNA adenine methyltransferase identification (DamID) method, in which a DNA-binding protein is fused to a bacterial methyltransferase. Adenines near binding sites in the genome are methylated by the fusion protein, allowing subsequent enrichment and sequencing. In practice, protein levels must be kept very low to prevent toxicity. Southall et al. achieved this by cloning the fusion protein downstream of a protein-coding region, which resulted in weak translational initiation. Their targeted DamID, or 'TaDa', method allows the use of inducible Gal4 drivers for specific cell types rather than constitutive basal promoters. The researchers profiled chromatin-binding proteins and RNA polymerase II to determine epigenetic status and gene expression in neural stem cells in the fruit fly.
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Following cell-type–specific gene expression. Nat Methods 10, 699 (2013). https://doi.org/10.1038/nmeth.2578
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DOI: https://doi.org/10.1038/nmeth.2578
