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Fig. 2 | BMC Research Notes

Fig. 2

From: enChIP systems using different CRISPR orthologues and epitope tags

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

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