Fig. 3: GgeAgo can efficiently cleave DNA and RNA mediated by 5’P-gDNA in Mn2+. | Communications Biology

Fig. 3: GgeAgo can efficiently cleave DNA and RNA mediated by 5’P-gDNA in Mn2+.

From: A programmable pAgo nuclease with high activity and specificity for efficient DNA and RNA manipulation

Fig. 3

a DNA-guided nucleic acid cleavage by GgeAgo with various divalent cation. The reactions were performed for 15 min (DNA Target) or 20 min (RNA Target) at 50 °C. b Kinetics of nucleic acid cleavage using 5’P-gDNA by GgeAgo measured with FAM-labeled target ssDNA or RNA at 50 °C. The data were fitted to a single-exponential equation, and the resulting kobs value is shown. c Binding of GgeAgo to 18 nt gDNAs by fluorescence polarization assay. GgeAgo binds 5’P-gDNA and 5’OH-gDNA with average KD values of 8.23 ± 1.47 and 11.14 ±  1.88 nM, respectively. Binding of GgeAgo-gDNA complex to DNA target (d) and RNA Target (e) by fluorescence polarization assay. GgeAgo_DM preloaded 5’P-gDNA or 5’OH-gDNA binds target DNA with average KD values of 8.69 ± 1.28 and 19.98 ± 2.29 nM, and RNA with 10.65 ± 1.11 and 32.21 ± 4.90 nM, respectively. Results are from three independent experiments. Error bars represent the SD.

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