Background

Assumptions

Establishment

Algorithm design

The results of off-target assessment

Guide Sequence + PAM	BioTech	MIT	CFD	Mismatch
TAGCACCTTAAGCTTACCGC+CGG	95	100	100	[0, 0, 0, 0, 1]
TAAGCTTAAGGTGCTAGCCA+AGG	93	100	100	[0, 0, 0, 0, 4]
CGTCCGCCGTGGCCTTCACA+GGG	74	99	99	[0, 0, 0, 0, 13]
GTCGGCCCCTGTGAAGGCCA+CGG	74	94	96	[0, 0, 0, 1, 32]
GTCGGCCGGCGGTAAGCTTA+AGG	71	99	99	[0, 0, 0, 2, 6]
GGCCCCTGTGAAGGCCACGG+CGG	71	89	96	[0, 0, 0, 2, 57]
CGCCTTGTGGTGTCGGCCGG+CGG	64	93	96	[0, 0, 0, 2, 33]
GCTGGCGTCGGCCCCTGTGA+AGG	62	87	94	[0, 0, 0, 8, 127]
GCGTCCGCCGTGGCCTTCAC+AGG	62	91	96	[0, 0, 0, 2, 54]
GGTGTACTGGGGCGCCGCCG+CGG	60	77	93	[0, 0, 0, 4, 97]
GTCCGCCGTGGCCTTCACAG+GGG	57	92	95	[0, 0, 0, 2, 48]
GGACGCGGGCGTGGTGTACT+GGG	57	90	96	[0, 0, 0, 6, 67]
GCTAGCCAAGGCGCTGGCGT+CGG	54	65	93	[0, 0, 0, 12, 134]
TAAGGTGCTAGCCAAGGCGC+TGG	48	84	94	[0, 0, 0, 2, 35]
GACGCGGGCGTGGTGTACTG+GGG	47	91	96	[0, 0, 1, 6, 52]
CGGACGCGGGCGTGGTGTAC+TGG	41	84	93	[0, 0, 0, 9, 83]
TGTGAAGGCCACGGCGGACG+CGG	41	77	90	[0, 0, 0, 8, 103]
AGGGGCCGACGCCAGCGCCT+TGG	38	66	88	[0, 0, 1, 10, 131]
GTGAAGGCCACGGCGGACGC+GGG	35	69	93	[0, 0, 1, 11, 103]
GGCCACGGCGGACGCGGGCG+TGG	32	19	48	[0, 0, 16, 172, 1352]
TACCGCCGGCCGACACCACA+AGG	31	93	99	[0, 0, 1, 2, 8]
CTGCAGGTCGGCGTCCGCGG+CGG	15	38	80	[0, 1, 3, 43, 456]
CACCACGCCCGCGTCCGCCG+TGG	11	21	78	[0, 3, 1, 41, 362]

BioTech	MIT	CFD	CFD	Mismatch
99	100	100	100	[0, 0, 0, 0, 1]
96	100	100	100	[0, 0, 0, 0, 4]
88	100	100	99	[0, 0, 0, 0, 13]
83	99	100	96	[0, 0, 0, 1, 32]
80	99	100	99	[0, 0, 0, 2, 6]
80	98	100	96	[0, 0, 0, 2, 57]
80	98	100	96	[0, 0, 0, 2, 33]
78	98	100	94	[0, 0, 0, 8, 127]
77	98	99	96	[0, 0, 0, 2, 54]
74	98	99	93	[0, 0, 0, 4, 97]

The calculation of the cleavage possibility

Mismatch_Num	Max	Min	Mean	Std
0	99.99%	99.99%	99.99%	0.00%
1	99.99%	96.30%	99.32%	0.87%
2	99.99%	1.16%	53.34%	37.03%
3	99.66%	0.00%	9.47%	21.37%
4	67.65%	0.00%	0.34%	2.45%