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. Author manuscript; available in PMC: 2021 Jul 7.
Published in final edited form as: J Am Chem Soc. 2020 Jul 28;142(32):13954–13965. doi: 10.1021/jacs.0c06541

Figure 7. Azide-tailed sgRNA 1’Az localizes on telomeric region.

Figure 7

(a) mESCs were treated with dCas9-eGFP alone or its RNP complex with unlabeled sgRNA 1’ (targeting telomere), azide-labeled sgRNA 1’Az (targeting telomere) and azide-labeled sgRNA 2Az (targeting eGFP: nontargeting control). The cells were imaged in DAPI and eGFP channels, and maximally projected Z-stacks are shown. First two rows: RNP complex of 1’ and 1’Az directs the localization to telomeric regions visualized as nuclear puncta. Third row: RNP complex of 2Az (nontargeting) and dCas9-eGFP nonspecifically distributes throughout the nucleus with no observable puncta. Last row: incubation with dCas9-eGFP alone resulted in aggregates, which were mostly localized outside the nucleus (scale bar, 10 μm). (b) Maximally projected Z-stack image showing nuclear puncta localized to telomeres in a single embryonic stem cell obtained using RNP complex of 1’Az and dCas9-eGFP (scale bar, 10 μm). The experiments were performed as n = 2 biological replicates.