Literature DB >> 29075654

Micro-chromatin Immunoprecipation (μChIP) Protocol for Real-time PCR Analysis of a Limited Amount of Cells.

Sébastien Gillotin1,2, François Guillemot1.   

Abstract

Chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) is an important strategy to study gene regulation. When availability of cells is limited, however, it can be useful to focus on specific genes to investigate in depth the role of transcription factors or histone marks. Unfortunately, performing ChIP experiments to study transcription factors' binding to DNA can be difficult when biological material is restricted. This protocol describes a robust method to perform μChIP for over-expressed or endogenous transcription factors using ~100,000 cells per ChIP experiment (Masserdotti et al., 2015). We also describe optimization steps, which we think are critical for this protocol to work and which can be used to further reduce the number of cells.

Entities:  

Year:  2016        PMID: 29075654      PMCID: PMC5654496          DOI: 10.21769/BioProtoc.1846

Source DB:  PubMed          Journal:  Bio Protoc        ISSN: 2331-8325


  2 in total

1.  Binding of c-Myc to chromatin mediates mitogen-induced acetylation of histone H4 and gene activation.

Authors:  S R Frank; M Schroeder; P Fernandez; S Taubert; B Amati
Journal:  Genes Dev       Date:  2001-08-15       Impact factor: 11.361

2.  Transcriptional Mechanisms of Proneural Factors and REST in Regulating Neuronal Reprogramming of Astrocytes.

Authors:  Giacomo Masserdotti; Sébastien Gillotin; Bernd Sutor; Daniela Drechsel; Martin Irmler; Helle F Jørgensen; Steffen Sass; Fabian J Theis; Johannes Beckers; Benedikt Berninger; François Guillemot; Magdalena Götz
Journal:  Cell Stem Cell       Date:  2015-06-25       Impact factor: 25.269

  2 in total
  2 in total

1.  Isolation of Chromatin-bound Proteins from Subcellular Fractions for Biochemical Analysis.

Authors:  Sébastien Gillotin
Journal:  Bio Protoc       Date:  2018-10-05

2.  Astrocytes and neurons share region-specific transcriptional signatures that confer regional identity to neuronal reprogramming.

Authors:  Álvaro Herrero-Navarro; Lorenzo Puche-Aroca; Verónica Moreno-Juan; Alejandro Sempere-Ferràndez; Ana Espinosa; Rafael Susín; Laia Torres-Masjoan; Eduardo Leyva-Díaz; Marisa Karow; María Figueres-Oñate; Laura López-Mascaraque; José P López-Atalaya; Benedikt Berninger; Guillermina López-Bendito
Journal:  Sci Adv       Date:  2021-04-07       Impact factor: 14.136

  2 in total

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