Supplementary Materials

CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity

Janice S. Chen, Enbo Ma, Lucas B. Harrington, Maria Da Costa, Xinran Tian, Joel M. Palefsky, Jennifer A. Doudna

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S15
  • Table S1
  • References
Correction (18 February 2021): After publication, a data analysis error was brought to the authors’ attention. The initial velocity (V0) plotted in Fig. 2D and fig. S7 was calculated incorrectly. The fluorescence intensity was used directly in the calculation of V0 rather than being converted first to molar concentrations. As a result, there is a ~100-fold difference between the reported and the correct Michaelis-Menten values. LbCas12a-crRNA bound to a ssDNA activator molecule catalyzed trans-ssDNA cleavage at a rate of ~3 turnovers per second with a catalytic efficiency (kcat/Km) of 5.0 × 106 s−1 M−1 rather than the reported ~250 turnovers per second and kcat/Km of 5.1 × 108 s−1 M−1. When bound to a dsDNA activator, LbCas12a-crRNA catalyzed ~17 turnovers per second with a kcat/Km of 1.7 × 10;7 s−1 M−1 (Fig. 2D and fig. S7), rather than the reported ~1250 turnovers per second and kcat/Km of 1.7 × 109 s−1 M−1. Fig. 2D, fig. S7, and the associated text have been corrected to describe the calculation of the Michaelis-Menten values, following the methods described in Cofsky et al. (2020), which has been added to the reference list. The authors thank J. Santiago for pointing out the error (
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