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. Author manuscript; available in PMC: 2017 Nov 29.
Published in final edited form as: ACS Synth Biol. 2017 Mar 20;6(6):1103–1113. doi: 10.1021/acssynbio.7b00050

Table 1.

Rate Constant of AceCas9 Cleaving Various Substrates

20-nt spacer sgRNA
substrate rate constant (min−1) S.D.a (min−1) goodness of fit (R square)
dsDNA + PAM 0.104 0.005 0.9852
T(−1)C bulge dsDNA + PAM 0.179 0.007 0.9904
T(−1)C bulge dsDNA − PAM 0.098 0.009 0.9349
ssDNA 0.65 0.10 0.8348
Supercoiled DNA plasmid (37 °C) 0.093 0.003 0.9950
Supercoiled DNA plasmid (50 °C) 0.26 0.02 0.9761
Gyrase-treated supercoiled DNA plasmid (50 °C) 0.30 0.02 0.9649
BamHI-treated linearized DNA plasmid (50 °C) 0.10 0.01 0.9484
24-nt spacer sgRNA
substrate rate constant (min−1) S.D.a (min−1) goodness of fit (R square)
Supercoiled DNA plasmid (37 °C) 0.28 0.01 0.9538
Supercoiled DNA plasmid (50 °C) 1.04 0.03 0.9684
BamHI-treated linearized DNA plasmid (50 °C) 0.53 0.05 0.9798
a

SD denotes standard deviation of experiments in triplicate.