Szczesnik, Tomasz and Ho, Joshua W. K. and Sherwood, Richard (2019) Dam mutants provide improved sensitivity and spatial resolution for profiling transcription factor binding. Epigenetics & Chromatin, 12 (1). ISSN 1756-8935
Full text not available from this repository.Abstract
DamID, in which a protein of interest is fused to Dam methylase, enables mapping of protein-DNA binding through readout of adenine methylation in genomic DNA. DamID offers a compelling alternative to chromatin immunoprecipitation sequencing (ChIP-Seq), particularly in cases where cell number or antibody availability is limiting. This comes at a cost, however, of high non-specific signal and a lowered spatial resolution of several kb, limiting its application to transcription factor-DNA binding. Here we show that mutations in Dam, when fused to the transcription factor Tcf7l2, greatly reduce non-specific methylation. Combined with a simplified DamID sequencing protocol, we find that these Dam mutants allow for accurate detection of transcription factor binding at a sensitivity and spatial resolution closely matching that seen in ChIP-seq.
| Item Type: | Article |
|---|---|
| Subjects: | R Medicine > R Medicine (General) |
| Depositing User: | Repository Administrator |
| Date Deposited: | 20 Nov 2019 00:16 |
| Last Modified: | 20 Nov 2019 00:16 |
| URI: | https://eprints.victorchang.edu.au/id/eprint/890 |
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