Table 2.

Cas12a2 collateral cleavage kinetics.

Substrate 10-nt Collateral Substrates	Km (μM)	Vmax (nM sec−1)	kcat (sec−1)	kcat/Km (M−1 sec−1)
ssRNA	0.09 ± 0.01	0.39 ± 0.01	0.0260 ± 0.0007	(2.9 ± 0.3) × 105
5'-UGUAACGACC-3'
Cas12a2 (Y465A)	0.104 ± 0.008	0.422 ± 0.009	0.0281 ± 0.0006	(2.7 ± 0.2) × 105
Cas12a2 (Y1080A)	0.12 ± 0.01	0.359 ± 0.009	0.0239 ± 0.0006	(2.0 ± 0.2) × 105
Cas12a2 (Y1069A)	0.16 ± 0.01	0.58 ± 0.01	0.0387 ± 0.0007	(2.4 ± 0.2) × 105
Cas12a2 (F1092A)	3.6 ± 3.7	0.5 ± 0.4	0.03 ± 0.03	(1.0 ± 1.7) × 104
ssDNA	0.025 ± 0.007	0.185 ± 0.004	0.0123 ± 0.0003	(4.9 ± 1.4) × 105
5'-TGTAACGACC-3'
Cas12a2 (Y465A)	0.025 ± 0.004	0.243 ± 0.005	0.0162 ± 0.0003	(6.5 ± 1.1) × 105
Cas12a2 (Y1080A)	0.015 ± 0.003	0.189 ± 0.004	0.0126 ± 0.0003	(8.4 ± 1.7) × 105
Cas12a2 (Y1069A)	0.030 ± 0.007	0.251 ± 0.009	0.0167 ± 0.0006	(5.6 ± 1.3) × 105
Cas12a2 (F1092A)	0.25 ± 0.06	0.054 ± 0.004	0.0036 ± 0.0003	(1.4 ± 0.4) × 104
dsDNA	0.042 ± 0.006	0.18 ± 0.005	0.0120 ± 0.0003	(2.9 ± 0.4) × 105
5'-F-TGTAACGACC-Q-3' 3'- ACATTGCTGG −5'
Cas12a2 (Y465A)	0.10 ± 0.03	0.18 ± 0.01	0.0120± 0.0007	(1.2 ± 0.4) × 105
Cas12a2 (Y1080A)	0.03 ± 0.01	0.14 ± 0.01	0.0093 ± 0.0007	(3.1 ± 1.1) × 105
Cas12a2 (Y1069A)	0.10 ± 0.02	0.19 ± 0.01	0.0127 ± 0.0007	(1.3 ± 0.3) × 105
Cas12a2 (F1092A)	--	--	--	--
dsDNA( Quencher on opposite strand)	0.040 ± 0.005	0.218 ± 0.005	0.0145 ± 0.0003	(3.6 ± 0.5) × 105
5'-F-TGTAACGACC −3' 3'- ACATTGCTGG-Q-5'