CATCH_IT
Covalent Attachment of Tags to Capture Histones and Identify Turnover
CATCH-IT is a direct method for measuring nucleosome turnover dynamics genome-wide (Deal et al., 2010). In this method, cells are treated briefly with the methionine surrogate azidohomoalanine (Aha), which couples biotin to nucleosomes containing newly incorporated histones. The labeled chromatin is affinity-purified with streptavidin, washed stringently to remove nonhistone proteins, and analyzed using tiling microarrays.
Advantages:
- Can determine differences in nucleosome turnover across the genome
Disadvantages:
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Zentner G. E. and Henikoff S. High-resolution digital profiling of the epigenome. Nat Rev Genet. 2014;15:814-827
References:
Perez-Lluch S., Blanco E., Tilgner H., et al. Absence of canonical marks of active chromatin in developmentally regulated genes. Nat Genet. 2015;47:1158-1167
Skene P. J., Hernandez A. E., Groudine M. and Henikoff S. The nucleosomal barrier to promoter escape by RNA polymerase II is overcome by the chromatin remodeler Chd1. Elife. 2014;3:e02042
Weber C. M., Ramachandran S. and Henikoff S. Nucleosomes are context-specific, H2A.Z-modulated barriers to RNA polymerase. Mol Cell. 2014;53:819-830
Teves S. S. and Henikoff S. Transcription-generated torsional stress destabilizes nucleosomes. Nat Struct Mol Biol. 2014;21:88-94
Yildirim O., Hung J. H., Cedeno R. J., Weng Z., Lengner C. J. and Rando O. J. A system for genome-wide histone variant dynamics in ES cells reveals dynamic MacroH2A2 replacement at promoters. PLoS Genet. 2014;10:e1004515
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History: CATCH_IT
Revision by sbrumpton on 2017-06-21 09:28:07 - Show/Hide
Covalent Attachment of Tags to Capture Histones and Identify Turnover
CATCH-IT is a direct method for measuring nucleosome turnover dynamics genome-wide (Deal et al., 2010). In this method, cells are treated briefly with the methionine surrogate azidohomoalanine (Aha), which couples biotin to nucleosomes containing newly incorporated histones. The labeled chromatin is affinity-purified with streptavidin, washed stringently to remove nonhistone proteins, and analyzed using tiling microarrays.
Advantages:- Can determine differences in nucleosome turnover across the genome
Disadvantages:Reagents:Illumina Library prep and Array Kit SelectorReviews:Zentner G. E. and Henikoff S. High-resolution digital profiling of the epigenome. Nat Rev Genet. 2014;15:814-827References:Perez-Lluch S., Blanco E., Tilgner H., et al. Absence of canonical marks of active chromatin in developmentally regulated genes. Nat Genet. 2015;47:1158-1167Skene P. J., Hernandez A. E., Groudine M. and Henikoff S. The nucleosomal barrier to promoter escape by RNA polymerase II is overcome by the chromatin remodeler Chd1. Elife. 2014;3:e02042Weber C. M., Ramachandran S. and Henikoff S. Nucleosomes are context-specific, H2A.Z-modulated barriers to RNA polymerase. Mol Cell. 2014;53:819-830Teves S. S. and Henikoff S. Transcription-generated torsional stress destabilizes nucleosomes. Nat Struct Mol Biol. 2014;21:88-94Yildirim O., Hung J. H., Cedeno R. J., Weng Z., Lengner C. J. and Rando O. J. A system for genome-wide histone variant dynamics in ES cells reveals dynamic MacroH2A2 replacement at promoters. PLoS Genet. 2014;10:e1004515