Table 1 Properties of DNA-guided TtAgo binding to DNA targets with different extents of complementarity to the guide-strand at various temperatures

From: An automated Bayesian pipeline for rapid analysis of single-molecule binding data

 

kon (M−1 s−1)

 

23°C

37°C

45°C

55°C

Fully complementary

6.5 ± 0.1 × 107

8.5 ± 0.1 × 107

12.0 ± 0.1 × 107

11.1 ± 0.1 × 107

Seed-matched

7.0 ± 0.1 × 107

6.9 ± 0.1 × 107

9.0 ± 0.1 × 107

12.0 ± 0.1 × 107

Seed-matched + 3′ sup. pairing

7.5 ± 0.1 × 107

5.5 ± 0.1 × 107

9.0 ± 0.1 × 107

13.5 ± 0.1 × 107

 

koff (s−1)

 

23°C

37°C

45°C

55°C

Fully complementary

« PB

« PB

« PB

« PB

Seed-matched

0.10 ± 0.01

0.61 ± 0.01

2.0 ± 0.1

18.0 ± 0.3

Seed-matched + 3′ sup. pairing

0.09 ± 0.01

0.67 ± 0.01

2.0 ± 0.1

13.2 ± 0.3

  1. Values of kon and koff were derived from data collected from several hundred individual DNA target molecules ( > 1100); standard error from bootstrapping is reported. Representative fluorescence intensity time traces and rastergrams summarizing traces of individual target molecules are shown in Supplementary Fig. 2
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