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. 2018 Feb 27;11:154. doi: 10.1186/s13104-018-3262-4

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© The Author(s) 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 2 — enChIP using the S. pyogenes CRISPR system tagged with the 2xAM-tag. a The S. pyogenes CRISPR system tagged with 2xAM-tag for enChIP. The system is composed of a fusion protein Sp-dCas9-2xAM (consisting of Sp-dCas9, an NLS, and a 2xAM-tag) and a gRNA. b Scheme of the enChIP system using S. pyogenes CRISPR tagged with the 2xAM-tag. After expression of Sp-dCas9-2xAM and a gRNA for locus-tagging in target cells, enChIP is performed using an Ab against the AM-tag, as shown in Fig. 1b. c Expression of Sp-dCas9-2xAM. pLenti_dCas9-2xAM or pLenti_dCas9-2xAM_hIRF-1 was transduced into HT1080 cells. After puromycin selection, expression of Sp-dCas9-2xAM was detected by immunoblot analysis with Ab against AM-tag. d Isolation of the IRF-1 locus by the S. pyogenes CRISPR system tagged with 2xAM-tag. Real-time PCR analysis was performed on chromatin complexes isolated by enChIP. An irrelevant locus (SOX2) was analysed as a negative control. The S. pyogenes CRISPR system tagged with the 3xFLAG-tag was used as a positive control for enChIP