grID Information

Summary

grID: hs025713225
Sequence (23-mer): GGAGCAGAGAGCGAAGCGGGAGG
grID Score: moderate (400)
Doench (2014) Score: 0.44736
Doench (2016) Score: 0.57389
Cas9: S. pyogenes

Genomic Context

Genomic Location: chr12:6643694-6643672
Strand: -
gRNA location: exon:refGene(NM_002046.exon0)

Nearest Genes

refseqgenechromstartenddistance
NM_002046 GAPDHchr12664358466475370
NM_001256799 GAPDHchr1266444086647537736
NM_014865 NCAPD2chr12660329766411322562
NM_001039670 IFFO1chr12664869366652495021
NM_001193457 IFFO1chr12664869366652495021

DNase I hypersensitive sites from ENCODE

DNase I hypersensitivity region:chr12:6643560-6643710
Numbers of Cell Types:123
Cell Types:8988t, A549, AG04449, AG04450, AG09309, AG09319, AG10803, Adult_CD4_Th0, AoAF, AoSMC, BE2_C, BJ, Caco-2, CD14+, CD20+, CD34+_Mobilized, CMK, Chorion, Cll, HSMM_emb, Fibrobl, Fibrop, GM06990, GM12864, GM12865, GM12878, Gliobla, Gm12891, Gm12892, Gm18507, Gm19238, Gm19239, Gm19240, H9es, HAEpiC, HAc, HA-h, HBMEC, HCF, HCFaa, HCM, HCPEpiC, HCT-116, HConF, HEEpiC, H1-hESC, HFF, HFF-Myc, HGF, HIPEpiC, HL-60, HMEC, HMF, HMVEC-LBl, HMVEC-LLy, HMVEC-dAd, HMVEC-dBl-Ad, HMVEC-dBl-Neo, HMVEC-dLy-Ad, HMVEC-dLy-Neo, HMVEC-dNeo, HNPCEpiC, HPAEC, HPAF, HPF, HPdLF, HRCEpiC, HRE, HRGEC, HRPEpiC, HSMM, HSMM_FSHD, HUVEC, HVMF, HeLa-S3, HeLa-S3s3Ifna4h, HepG2, Hepatocytes, HPDE6-E6E7, HTR8svn, Huh-7, Huh-7.5, Ips, Ishikawa, Ishikawa, Jurkat, K562, LNCap, LNCaP, MCF-7, MCF-7, Medullo, Melano, Myometr, NB4, NH-A, NHDF-Ad, NHDF-neo, NHEK, NHLF, NT2-D1, Osteobl, PanIsletD, Panislets, Phte, PrEC, Progfib, RPTEC, RWPE1, SAEC, SK-N-SH, SK-N-MC, SkMC, Stellate, T-47D, Urothelia, UrotheliaUt189, WERI-Rb-1, WI-38, WI-38_TAM, H1-hESC, Th1, Th2,

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Sequence Analysis

Target Sequence (RNA): GGAGCAGAGAGCGAAGCGGG
Target Sequence Occurrence: unique
PAM motif: AGG
Homopolymeric Runs: none
Pol III Terminator: none

Thermodynamic Properties

GC-content (%): 70
GC-score: high
Tm (°C): 68.13
Tm score: moderate
RNAfold (dot bracket): ...((.....))........
deltaG (Kcal/mol): -0.50
Predicted Folding: very low secondary structure

Offtarget Analysis

Total Predicted Genomic Targets: 38
Total Genomic Sites With 0 Mismatches: 1
Total Genomic Sites With 1 Mismatches:
Total Genomic Sites With 2 Mismatches: 3
Total Genomic Sites With 3 Mismatches: 34
1chr12:6643671-66436940
CCTCCCGCTTCGCTCTCTGCTCC
|||||||||||||||||||||||
CCTCCCGCTTCGCTCTCTGCTCC
2chr16:1630456-16304792
CCTCCCGCTTCGCTCTCTGCTCC
||||||||||||||||||**|||
CCTCCCGCTTCGCTCTCTCGTCC
3chr8:144272925-1442729482
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||||||||||*|||
GGAGGAGAGAGCGAAGCGGAAGG
4chr8:125526502-1255265252
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*|||*|||||||||||
GGAGCAGGGAGGGAAGCGGGAGG
5chr4:126512861-1265128843
GGAGCAGAGAGCGAAGCGGGAGG
|||**|*||||||||||||||||
GGAAGACAGAGCGAAGCGGGAGG
6chr16:75822470-758224933
CCTCCCGCTTCGCTCTCTGCTCC
||||||||||||||||||*|**|
CCTCCCGCTTCGCTCTCTCCAGC
7chr22:40232813-402328363
CCTCCCGCTTCGCTCTCTGCTCC
||||||||||||||||||*|**|
CCTCCCGCTTCGCTCTCTCCAGC
8chr7:100281024-1002810473
CCTCCCGCTTCGCTCTCTGCTCC
|||||||||||||*||**|||||
CCTCCCGCTTCGCACTGAGCTCC
9chr1:241564676-2415646993
GGAGCAGAGAGCGAAGCGGGAGG
|**|*||||||||||||||||||
GAGGAAGAGAGCGAAGCGGGAGG
10chr4:70540963-705409863
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||*||||||||||
GGAAGAGAGAGCAAAGCGGGAGG
11chr3:186910288-1869103113
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||||||*||||||
GGAAGAGAGAGCGAAGGGGGAGG
12chr1:6784296-67843193
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||||||*||||||
GGAAAAGAGAGCGAAGGGGGAGG
13chr6:161402844-1614028673
CCTCCCGCTTCGCTCTCTGCTCC
||||||*|||||||||||**|||
CCTCCCCCTTCGCTCTCTCGTCC
14chr5:122976932-1229769553
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||*||||||||||
GGAAGAGAGAGCAAAGCGGGAGG
15chr5:137802419-1378024423
CCTCCCGCTTCGCTCTCTGCTCC
|||||||||||*||||||*||*|
CCTCCCGCTTCCCTCTCTCCTGC
16chr17:34901384-349014073
CCTCCCGCTTCGCTCTCTGCTCC
|||||||||*|||||*||||*||
CCTCCCGCTACGCTCACTGCACC
17chr2:241346735-2413467583
CCTCCCGCTTCGCTCTCTGCTCC
|||||||||||*||||||**|||
CCTCCCGCTTCCCTCTCTCGTCC
18chr17:71467268-714672913
CCTCCCGCTTCGCTCTCTGCTCC
||||||*||||*||||||||*||
CCTCCCCCTTCCCTCTCTGCACC
19chr18:56972450-569724733
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||*||||||*|||||
GGAGCAGGGAACGAAGCAGGAGG
20chr5:172074238-1720742613
GGAGCAGAGAGCGAAGCGGGAGG
*||||||||||*||||*||||||
AGAGCAGAGAGGGAAGAGGGAGG
21chr11:118767688-1187677113
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||||*|||*||||||
GGAGCAGGGAGCCAAGGGGGAGG
22chr10:115500475-1155004983
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||||||*|*||||||
GGAGCAGGGAGCGAGGTGGGAGG
23chr19:34165551-341655743
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*|||*|*|||||||||
GGAGCAGGGAGAGGAGCGGGAGG
24chr10:104503676-1045036993
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||***|||||||||||
GGAGCAGAGCCGGAAGCGGGAGG
25chr19:1790197-17902203
CCTCCCGCTTCGCTCTCTGCTCC
||||||*||||*||||||*||||
CCTCCCCCTTCCCTCTCTCCTCC
26chrX:25447138-254471613
CCTCCCGCTTCGCTCTCTGCTCC
||||||*||||*||||||*||||
CCTCCCCCTTCCCTCTCTCCTCC
27chr19:8018195-80182183
CCTCCCGCTTCGCTCTCTGCTCC
||||||*||||*||||||*||||
CCTCCCCCTTCCCTCTCTCCTCC
28chr20:12613995-126140183
CCTCCCGCTTCGCTCTCTGCTCC
||||||*||||*||||||*||||
CCTCCCCCTTCCCTCTCTCCTCC
29chrX:49524184-495242073
GGAGCAGAGAGCGAAGCGGGAGG
|||*||||||||||||*||||*|
GGAACAGAGAGCGAAGGGGGAAG
30chr5:89676139-896761623
CCTCCCGCTTCGCTCTCTGCTCC
|||*|||||||*||||||*||||
CCTGCCGCTTCCCTCTCTCCTCC
31chr16:79781007-797810303
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGAAGAGAGAGAAGAGGGAGG
32chr9:38691218-386912413
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||||**|||||
GGAGCAGAGAGAGAAGGAGGAGG
33chr11:57009805-570098283
CCTCCCGCTTCGCTCTCTGCTCC
||||||*|*|*||||||||||||
CCTCCCCCATGGCTCTCTGCTCC
34chr7:28199075-281990983
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||**|*||||||||
GGAGCAGAGAGGAAGGCGGGAGG
35chr10:6078396-60784193
CCTCCCGCTTCGCTCTCTGCTCC
|||*||||*||*|||||||||||
CCTGCCGCATCCCTCTCTGCTCC
36chr19:3551831-35518543
CCTCCCGCTTCGCTCTCTGCTCC
*|||||*||||*|||||||||||
GCTCCCCCTTCCCTCTCTGCTCC
37chr3:11174765-111747883
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||||||||**|
GGAGCAGAGAGTGAAGCGGGGAG
38chr6:160390328-1603903513
CCTCCCGCTTCGCTCTCTGCTCC
||*|||||**|||||||||||||
CCACCCGCAACGCTCTCTGCTCC
1
Number of Mismatches:0
Location:chr12:6643671-6643694
Genomic Sequence:GGAGCAGAGAGCGAAGCGGGAGG
Strand:-
Genomic Region:exon(NM_002046.exon0)
3' PAM Orientation:GGAGCAGAGAGCGAAGCGGGAGG
Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||||||||||||||
GGAGCAGAGAGCGAAGCGGGAGG
Mismatch: 


2


Number of Mismatches:2


Location:chr16:1630456-1630479


Genomic Sequence:GGACGAGAGAGCGAAGCGGGAGG


Strand:-


Genomic Region:gene(IFT140)



3' PAM Orientation:GGACGAGAGAGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**||||||||||||||||||
GGACGAGAGAGCGAAGCGGGAGG


Mismatch:

...C...................
....G..................
Distal mismatch
Distal mismatch


3


Number of Mismatches:2


Location:chr8:144272925-144272948


Genomic Sequence:GGAGGAGAGAGCGAAGCGGAAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGGAGAGAGCGAAGCGGAAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||||||||||*|||
GGAGGAGAGAGCGAAGCGGAAGG


Mismatch:

....G..................
...................A...
Distal mismatch
Proximal seed mismatch


4


Number of Mismatches:2


Location:chr8:125526502-125526525


Genomic Sequence:GGAGCAGGGAGGGAAGCGGGAGG


Strand:+


Genomic Region:gene(TATDN1)



3' PAM Orientation:GGAGCAGGGAGGGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*|||*|||||||||||
GGAGCAGGGAGGGAAGCGGGAGG


Mismatch:

.......G...............
...........G...........
Distal seed mismatch
Proximal seed mismatch


5


Number of Mismatches:3


Location:chr4:126512861-126512884


Genomic Sequence:GGAAGACAGAGCGAAGCGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAAGACAGAGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|*||||||||||||||||
GGAAGACAGAGCGAAGCGGGAGG


Mismatch:

...A...................
....G..................
......C................
Distal mismatch
Distal mismatch
Distal mismatch


6


Number of Mismatches:3


Location:chr16:75822470-75822493


Genomic Sequence:GCTGGAGAGAGCGAAGCGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GCTGGAGAGAGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|**|*||||||||||||||||||
GCTGGAGAGAGCGAAGCGGGAGG


Mismatch:

.C.....................
..T....................
....G..................
Distal mismatch
Distal mismatch
Distal mismatch


7


Number of Mismatches:3


Location:chr22:40232813-40232836


Genomic Sequence:GCTGGAGAGAGCGAAGCGGGAGG


Strand:-


Genomic Region:gene(ENTHD1)



3' PAM Orientation:GCTGGAGAGAGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|**|*||||||||||||||||||
GCTGGAGAGAGCGAAGCGGGAGG


Mismatch:

.C.....................
..T....................
....G..................
Distal mismatch
Distal mismatch
Distal mismatch


8


Number of Mismatches:3


Location:chr7:100281024-100281047


Genomic Sequence:GGAGCTCAGTGCGAAGCGGGAGG


Strand:-


Genomic Region:exon(NM_022574.exon17)



3' PAM Orientation:GGAGCTCAGTGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||**||*|||||||||||||
GGAGCTCAGTGCGAAGCGGGAGG


Mismatch:

.....T.................
......C................
.........T.............
Distal mismatch
Distal mismatch
Distal seed mismatch


9


Number of Mismatches:3


Location:chr1:241564676-241564699


Genomic Sequence:GAGGAAGAGAGCGAAGCGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GAGGAAGAGAGCGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|**|*||||||||||||||||||
GAGGAAGAGAGCGAAGCGGGAGG


Mismatch:

.A.....................
..G....................
....A..................
Distal mismatch
Distal mismatch
Distal mismatch


10


Number of Mismatches:3


Location:chr4:70540963-70540986


Genomic Sequence:GGAAGAGAGAGCAAAGCGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAAGAGAGAGCAAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||*||||||||||
GGAAGAGAGAGCAAAGCGGGAGG


Mismatch:

...A...................
....G..................
............A..........
Distal mismatch
Distal mismatch
Proximal seed mismatch


11


Number of Mismatches:3


Location:chr3:186910288-186910311


Genomic Sequence:GGAAGAGAGAGCGAAGGGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAAGAGAGAGCGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||||||*||||||
GGAAGAGAGAGCGAAGGGGGAGG


Mismatch:

...A...................
....G..................
................G......
Distal mismatch
Distal mismatch
Proximal seed mismatch


12


Number of Mismatches:3


Location:chr1:6784296-6784319


Genomic Sequence:GGAAAAGAGAGCGAAGGGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAAAAGAGAGCGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||||||*||||||
GGAAAAGAGAGCGAAGGGGGAGG


Mismatch:

...A...................
....A..................
................G......
Distal mismatch
Distal mismatch
Proximal seed mismatch


13


Number of Mismatches:3


Location:chr6:161402844-161402867


Genomic Sequence:GGACGAGAGAGCGAAGGGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGACGAGAGAGCGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||||||*||||||
GGACGAGAGAGCGAAGGGGGAGG


Mismatch:

...C...................
....G..................
................G......
Distal mismatch
Distal mismatch
Proximal seed mismatch


14


Number of Mismatches:3


Location:chr5:122976932-122976955


Genomic Sequence:GGAAGAGAGAGCAAAGCGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAAGAGAGAGCAAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**|||||||*||||||||||
GGAAGAGAGAGCAAAGCGGGAGG


Mismatch:

...A...................
....G..................
............A..........
Distal mismatch
Distal mismatch
Proximal seed mismatch


15


Number of Mismatches:3


Location:chr5:137802419-137802442


Genomic Sequence:GCAGGAGAGAGGGAAGCGGGAGG


Strand:-


Genomic Region:gene(EGR1)



3' PAM Orientation:GCAGGAGAGAGGGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|*||*||||||*|||||||||||
GCAGGAGAGAGGGAAGCGGGAGG


Mismatch:

.C.....................
....G..................
...........G...........
Distal mismatch
Distal mismatch
Proximal seed mismatch


16


Number of Mismatches:3


Location:chr17:34901384-34901407


Genomic Sequence:GGTGCAGTGAGCGTAGCGGGAGG


Strand:-


Genomic Region:gene(GGNBP2)



3' PAM Orientation:GGTGCAGTGAGCGTAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||*||||*|||||*|||||||||
GGTGCAGTGAGCGTAGCGGGAGG


Mismatch:

..T....................
.......T...............
.............T.........
Distal mismatch
Distal seed mismatch
Proximal seed mismatch


17


Number of Mismatches:3


Location:chr2:241346735-241346758


Genomic Sequence:GGACGAGAGAGGGAAGCGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGACGAGAGAGGGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||**||||||*|||||||||||
GGACGAGAGAGGGAAGCGGGAGG


Mismatch:

...C...................
....G..................
...........G...........
Distal mismatch
Distal mismatch
Proximal seed mismatch


18


Number of Mismatches:3


Location:chr17:71467268-71467291


Genomic Sequence:GGTGCAGAGAGGGAAGGGGGAGG


Strand:-


Genomic Region:gene(SDK2)



3' PAM Orientation:GGTGCAGAGAGGGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||*||||||||*||||*||||||
GGTGCAGAGAGGGAAGGGGGAGG


Mismatch:

..T....................
...........G...........
................G......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


19


Number of Mismatches:3


Location:chr18:56972450-56972473


Genomic Sequence:GGAGCAGGGAACGAAGCAGGAGG


Strand:+


Genomic Region:gene(CPLX4)



3' PAM Orientation:GGAGCAGGGAACGAAGCAGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||*||||||*|||||
GGAGCAGGGAACGAAGCAGGAGG


Mismatch:

.......G...............
..........A............
.................A.....
Distal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


20


Number of Mismatches:3


Location:chr5:172074238-172074261


Genomic Sequence:AGAGCAGAGAGGGAAGAGGGAGG


Strand:+


Genomic Region:gene(NEURL1B)



3' PAM Orientation:AGAGCAGAGAGGGAAGAGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
*||||||||||*||||*||||||
AGAGCAGAGAGGGAAGAGGGAGG


Mismatch:

A......................
...........G...........
................A......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


21


Number of Mismatches:3


Location:chr11:118767688-118767711


Genomic Sequence:GGAGCAGGGAGCCAAGGGGGAGG


Strand:+


Genomic Region:three(NM_182557.utr7)



3' PAM Orientation:GGAGCAGGGAGCCAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||||*|||*||||||
GGAGCAGGGAGCCAAGGGGGAGG


Mismatch:

.......G...............
............C..........
................G......
Distal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


22


Number of Mismatches:3


Location:chr10:115500475-115500498


Genomic Sequence:GGAGCAGGGAGCGAGGTGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGCAGGGAGCGAGGTGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*||||||*|*||||||
GGAGCAGGGAGCGAGGTGGGAGG


Mismatch:

.......G...............
..............G........
................T......
Distal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


23


Number of Mismatches:3


Location:chr19:34165551-34165574


Genomic Sequence:GGAGCAGGGAGAGGAGCGGGAGG


Strand:+


Genomic Region:gene(CHST8)



3' PAM Orientation:GGAGCAGGGAGAGGAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||*|||*|*|||||||||
GGAGCAGGGAGAGGAGCGGGAGG


Mismatch:

.......G...............
...........A...........
.............G.........
Distal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


24


Number of Mismatches:3


Location:chr10:104503676-104503699


Genomic Sequence:GGAGCAGAGCCGGAAGCGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGCAGAGCCGGAAGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||***|||||||||||
GGAGCAGAGCCGGAAGCGGGAGG


Mismatch:

.........C.............
..........C............
...........G...........
Distal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


25


Number of Mismatches:3


Location:chr19:1790197-1790220


Genomic Sequence:GGAGGAGAGAGGGAAGGGGGAGG


Strand:-


Genomic Region:gene(ATP8B3)



3' PAM Orientation:GGAGGAGAGAGGGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGGAGAGAGGGAAGGGGGAGG


Mismatch:

....G..................
...........G...........
................G......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


26


Number of Mismatches:3


Location:chrX:25447138-25447161


Genomic Sequence:GGAGGAGAGAGGGAAGGGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGAGGAGAGAGGGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGGAGAGAGGGAAGGGGGAGG


Mismatch:

....G..................
...........G...........
................G......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


27


Number of Mismatches:3


Location:chr19:8018195-8018218


Genomic Sequence:GGAGGAGAGAGGGAAGGGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGAGGAGAGAGGGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGGAGAGAGGGAAGGGGGAGG


Mismatch:

....G..................
...........G...........
................G......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


28


Number of Mismatches:3


Location:chr20:12613995-12614018


Genomic Sequence:GGAGGAGAGAGGGAAGGGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGAGGAGAGAGGGAAGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGGAGAGAGGGAAGGGGGAGG


Mismatch:

....G..................
...........G...........
................G......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


29


Number of Mismatches:3


Location:chrX:49524184-49524207


Genomic Sequence:GGAACAGAGAGCGAAGGGGGAAG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAACAGAGAGCGAAGGGGGAAG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||*||||||||||||*||||*|
GGAACAGAGAGCGAAGGGGGAAG


Mismatch:

...A...................
................G......
.....................A.
Distal mismatch
Proximal seed mismatch
PAM mismatch


30


Number of Mismatches:3


Location:chr5:89676139-89676162


Genomic Sequence:GGAGGAGAGAGGGAAGCGGCAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGAGGAGAGAGGGAAGCGGCAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*|||||||*|||
GGAGGAGAGAGGGAAGCGGCAGG


Mismatch:

....G..................
...........G...........
...................C...
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


31


Number of Mismatches:3


Location:chr16:79781007-79781030


Genomic Sequence:GGAGAAGAGAGAGAAGAGGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGAAGAGAGAGAAGAGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||*||||||*||||*||||||
GGAGAAGAGAGAGAAGAGGGAGG


Mismatch:

....A..................
...........A...........
................A......
Distal mismatch
Proximal seed mismatch
Proximal seed mismatch


32


Number of Mismatches:3


Location:chr9:38691218-38691241


Genomic Sequence:GGAGCAGAGAGAGAAGGAGGAGG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGCAGAGAGAGAAGGAGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||||**|||||
GGAGCAGAGAGAGAAGGAGGAGG


Mismatch:

...........A...........
................G......
.................A.....
Proximal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


33


Number of Mismatches:3


Location:chr11:57009805-57009828


Genomic Sequence:GGAGCAGAGAGCCATGGGGGAGG


Strand:-


Genomic Region:intergenic



3' PAM Orientation:GGAGCAGAGAGCCATGGGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
||||||||||||*|*|*||||||
GGAGCAGAGAGCCATGGGGGAGG


Mismatch:

............C..........
..............T........
................G......
Proximal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


34


Number of Mismatches:3


Location:chr7:28199075-28199098


Genomic Sequence:GGAGCAGAGAGGAAGGCGGGAGG


Strand:+


Genomic Region:gene(JAZF1)



3' PAM Orientation:GGAGCAGAGAGGAAGGCGGGAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||**|*||||||||
GGAGCAGAGAGGAAGGCGGGAGG


Mismatch:

...........G...........
............A..........
..............G........
Proximal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


35


Number of Mismatches:3


Location:chr10:6078396-6078419


Genomic Sequence:GGAGCAGAGAGGGATGCGGCAGG


Strand:-


Genomic Region:gene(IL2RA)



3' PAM Orientation:GGAGCAGAGAGGGATGCGGCAGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||*||||*|||
GGAGCAGAGAGGGATGCGGCAGG


Mismatch:

...........G...........
..............T........
...................C...
Proximal seed mismatch
Proximal seed mismatch
Proximal seed mismatch


36


Number of Mismatches:3


Location:chr19:3551831-3551854


Genomic Sequence:GGAGCAGAGAGGGAAGGGGGAGC


Strand:-


Genomic Region:gene(MFSD12)



3' PAM Orientation:GGAGCAGAGAGGGAAGGGGGAGC


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||||*|||||*
GGAGCAGAGAGGGAAGGGGGAGC


Mismatch:

...........G...........
................G......
......................C
Proximal seed mismatch
Proximal seed mismatch
PAM mismatch


37


Number of Mismatches:3


Location:chr3:11174765-11174788


Genomic Sequence:GGAGCAGAGAGTGAAGCGGGGAG


Strand:+


Genomic Region:intergenic



3' PAM Orientation:GGAGCAGAGAGTGAAGCGGGGAG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||*||||||||**|
GGAGCAGAGAGTGAAGCGGGGAG


Mismatch:

...........T...........
....................G..
.....................A.
Proximal seed mismatch
PAM N Offtarget
PAM mismatch


38


Number of Mismatches:3


Location:chr6:160390328-160390351


Genomic Sequence:GGAGCAGAGAGCGTTGCGGGTGG


Strand:-


Genomic Region:exon(NM_000876.exon0)



3' PAM Orientation:GGAGCAGAGAGCGTTGCGGGTGG


Alignment:
GGAGCAGAGAGCGAAGCGGGAGG
|||||||||||||**|||||*||
GGAGCAGAGAGCGTTGCGGGTGG


Mismatch:

.............T.........
..............T........
....................T..
Proximal seed mismatch
Proximal seed mismatch
PAM N Offtarget











Paired Nickase
Cas9(D10A)



	
PairOffsetOverhang

		
hs0257132282054

		
hs0257132312963

		
hs0257132323064

		
hs0257132333367

		
hs0257132354579

		
hs0257132364680

		
hs0257132374983

		
hs0257132385488

		
hs0257132395589

		
hs0257132405993

		
hs02571324267101

		
hs02571324370104

		
hs02571324474108

		
hs02571324575109

		
hs02571324676110

		
hs02571324779113

		
hs02571324880114

		
hs02571325186120

		
hs02571325290124




Sequence



Surrounding genomic sequence (500bp upstream and downstream target site)


Note: Repeats and low complexity DNA is shown in lower case; non repeating sequence is shown in upper case.







	















Screening Analysis


Restriction EnzymeRecognition SiteMatchSequence


Psp0357IIGCGAAGGCGAAGGGAGCAGAGAGCGAAGCGGGAGG


SpoDIGCGGRAGGCGGGAGGGAGCAGAGAGCGAAGCGGGAGG


Psp10HIIGRAGCAGGGAGCAGGGAGCAGAGAGCGAAGCGGGAGG


Cla11845IIIGCGAAGCGAAGGAGCAGAGAGCGAAGCGGGAGG


RlaIVCWGCAGGAGCAGAGAGCGAAGCGGGAGG




SNPs and Variation


Notes

	

        	×
        	Warning!  This gRNA contains a dinucleotide repeat sequence which can lead to greater off-targets.
    	




	

        	×
        	Warning!  This gRNA has a high GC content which can lead to greater off-targets.
    	



References



Jaskula-Ranga, V and Zack, DJ. 2016.  grID: A CRISPR-Cas9 guide RNA Database and Resource for Gene-Editing.
bioRxiv 097352; doi: https://doi.org/10.1101/097352


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