2016
DOI: 10.1016/j.molcel.2016.09.019
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Expanding the Circuitry of Pluripotency by Selective Isolation of Chromatin-Associated Proteins

Abstract: SummaryMaintenance of pluripotency is regulated by a network of transcription factors coordinated by Oct4, Sox2, and Nanog (OSN), yet a systematic investigation of the composition and dynamics of the OSN protein network specifically on chromatin is still missing. Here we have developed a method combining ChIP with selective isolation of chromatin-associated proteins (SICAP) followed by mass spectrometry to identify chromatin-bound partners of a protein of interest. ChIP-SICAP in mouse embryonic stem cells (ESC… Show more

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Cited by 89 publications
(121 citation statements)
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“…Downregulation of TRIM24 expression in human ES cells leads to spontaneous differentiation, which is in contrast to human breast cancer cells where TRIM24 depletion induces spontaneous apoptosis . Moreover, TRIM24 converges with Oct‐3/4, Sox2, and NANOG on multiple enhancers and suppresses the expression of developmental genes to support the pluripotent state of stem cells …”
Section: The Engagement Of Specific Trims In Stem Cell Maintenancementioning
confidence: 99%
“…Downregulation of TRIM24 expression in human ES cells leads to spontaneous differentiation, which is in contrast to human breast cancer cells where TRIM24 depletion induces spontaneous apoptosis . Moreover, TRIM24 converges with Oct‐3/4, Sox2, and NANOG on multiple enhancers and suppresses the expression of developmental genes to support the pluripotent state of stem cells …”
Section: The Engagement Of Specific Trims In Stem Cell Maintenancementioning
confidence: 99%
“…Overall, this study by Rafiee et al () presents a very useful improvement in the methodological toolbox available for the selective purification and identification of protein complexes and protein interaction networks associated with chromatin. The ChIP‐SICAP method offers several specific advantages: (i) it allows selective and reproducible isolation of chromatin‐associated proteins, together with the cognate, bound DNA element, (ii) it offers the possibility to distinguish between protein interactions taking place either, on or off, chromatin, and (iii) it facilitates combining proteomics analysis and ChIP‐seq in a single experiment.…”
Section: Sicap Methodsmentioning
confidence: 97%
“…In their recent work, Rafiee et al () describe a new method for the selective isolation of chromatin‐associated proteins (SICAP), which improves selectivity. The method involves ChIP of DNA‐protein complexes, combined with biotin tagging of DNA, to allow the specific isolation of DNA‐bound proteins on streptavidin beads, followed by MS‐based protein identification (Fig ).…”
Section: Sicap Methodsmentioning
confidence: 99%
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“…Crosslinking allows for the preservation of weak and transient interactions during harsh steps, such as sonication and stringent washing and has been optimized for different applications [16, 17, 18, 19]. From this, numerous methods have been developed that allow for proteomic analysis at specific genomic regions.…”
Section: Capturing Chromatin Proteins For Proteomic Analysismentioning
confidence: 99%