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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 8. Site-directed functionalization of telomeres by sgR-CLK was confirmed by ChIP-qPCR.

Figure 8

(a) Schematic diagram illustrating the steps involved in chromatin capture by in situ click reaction performed on the target-bound ternary complex using biotin-alkynes. While CuAAC reaction was performed using a biotin substrate containing a terminal alkyne, SPAAC reaction was performed using biotin-conjugated to a strained alkyne (sDIBO). See Figure S9 for complete structure of alkyne substrates. Biotinylated chromatin was enriched using streptavidin beads and subjected to qPCR analysis. (b) qPCR analysis of enriched chromatin obtained by CuAAC and SPAAC reactions using telomere-targeting sgRNA 1’Az and control nontargeting sgRNA 2Az. The enrichment is expressed relative to respective inputs after click reaction step. (c) A plot showing the fold enrichment of telomere DNA using sgRNA 1’Az normalized over sgRNA 2Az for CuAAC and SPAAC reactions (see Experimental Section for details). Values in b and c are denoted as mean ± s.d. performed as n = 3 technical replicates.