Table 2. Apparent dissociation constants for dCas9 with different guide RNA variants from the 989 bp ‘engineered’ DNA substrates (e.g. Figures 1D, 2C and D) that contain a series of fully- and partially- complementary protospacer sites.

Guide RNA variant	Overall dissociation constant between dCas9 and the engineered DNA substrate (±95% confidence)	Protospacer-specific dissociation constant for dCas9 and the full target on the engineered substrate (±95% confidence)
sgRNAa	2.70 nM (±1.58 nM)	44.67 nM (±1.04 nM)
tru-gRNAb	17.89 nM (±0.45 nM)	136.4 nM (±2.30 nM)
hp6-gRNAc	16.61 nM (±0.40 nM)	164.4 nM (±13.63 nM)
hp10-gRNAd	35.84 nM (±0.63 nM)	164.8 nM (±15.60 nM)

^aFull-length single-guide RNA (sgRNA).

^bTruncated sgRNA (first two nt at 5′- truncated).

^csgRNA with additional 5′- hairpin which overlaps six PAM-distal targeting nts (see text).

^dsgRNA with additional 5′- hairpin which overlaps ten PAM-distal targeting nts (see text).