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RNA-DamID reveals cell-type-specific binding of roX RNAs at chromatin-entry sites
Cheetham SW and Brand AH (2018) Nature Structural and Molecular Biology

RNA DamID detects interactions between long non-coding RNAs (lncRNAs) and the genome in a cell-type-specific manner in vivo. The technique is an adaptation of Targeted DamID and uses the MCP-MS2 system to bring lncRNAs and the Dam enzyme together.

The lncRNA of interest is tagged with three MS2 RNA stem-loops and the MS2 coat protein (MCP) is fused to the Dam protein. The Dam-MCP fusion is able to recognise the MS2-tagged lncRNA and will methylate the genome at sites of lncRNA binding or recruitment.

RNA-DamID produces comparable results to those obtained with ChIRP-Seq but is at least 75-fold more sensitive and has higher accuracy.

Available reagents (require MTA):

Please contact Andrea Brand

  • pUAST-attB-LT3-DAM-NLS-MCPx2 plasmid plasmid map

  • pUAST-attB-3xMS2 plasmid plasmid map

  • pUAST-attB-LT3-Dam-MSL3 plasmid plasmid map

  • pUAST-attB-3xMS2-rox2 plasmid plasmid map

  • UAS-LT3-Dam-NLS-MCPx2 Drosophila line

  • UAS-3xMS2-rox2 Drosophila line