Dam(ID)-profiling in B cells. Homo sapiens

Both chromatin access, and binding of AID in B cells are of major scientific interest. Therefore DamID was setup in B cells. AID specific binding could not be identified. In contrast to binding, DAM profiling provide a upmost useful tool in defining chromatin access in vivo. Overall design: Ramos B cells were transduced with Dam-only or with AID-Dam lentivirus. Within the cell, Dam methylates adenines near sites where the protein of interest would interacts (here AID) with the DNA. These methylated sequences are subsequently amplified by a methylation-specific PCR protocol as previsously described by Vogel et al (2007), and identified by hybridization to 2.1 Economy Whole Genome Nimblegen microarrays.