grID Information

Summary

grID: hs025713218
Sequence (23-mer): ATTTATAGAAACCGGGGGCGCGG
grID Score: high (800)
Doench (2014) Score: 0.49278
Doench (2016) Score: 0.53248
Cas9: S. pyogenes

Genomic Context

Genomic Location: chr12:6643660-6643638
Strand: -
gRNA location: exon:refGene(NM_002046.exon0)

Nearest Genes

refseqgenechromstartenddistance
NM_002046 GAPDHchr12664358466475370
NM_001256799 GAPDHchr1266444086647537770
NM_014865 NCAPD2chr12660329766411322528
NM_001039670 IFFO1chr12664869366652495055
NM_001193457 IFFO1chr12664869366652495055

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): AUUUAUAGAAACCGGGGGCG
Target Sequence Occurrence: unique
PAM motif: CGG
Homopolymeric Runs: yes
Pol III Terminator: none

Thermodynamic Properties

GC-content (%): 50
GC-score: moderate
Tm (°C): 51.54
Tm score: low
RNAfold (dot bracket): ....................
deltaG (Kcal/mol): 0.00
Predicted Folding: very low secondary structure

Offtarget Analysis

Total Predicted Genomic Targets: 1
Total Genomic Sites With 0 Mismatches: 1
Total Genomic Sites With 1 Mismatches:
Total Genomic Sites With 2 Mismatches:
Total Genomic Sites With 3 Mismatches:
1chr12:6643637-66436600
CCGCGCCCCCGGTTTCTATAAAT
|||||||||||||||||||||||
CCGCGCCCCCGGTTTCTATAAAT
1
Number of Mismatches:0
Location:chr12:6643637-6643660
Genomic Sequence:ATTTATAGAAACCGGGGGCGCGG
Strand:-
Genomic Region:exon(NM_002046.exon0)
3' PAM Orientation:ATTTATAGAAACCGGGGGCGCGG
Alignment:
ATTTATAGAAACCGGGGGCGCGG
|||||||||||||||||||||||
ATTTATAGAAACCGGGGGCGCGG
Mismatch:

Paired Nickase

Cas9(D10A)
PairOffsetOverhang
hs0257132285488
hs0257132316397
hs0257132326498
hs02571323367101
hs02571323579113
hs02571323680114
hs02571323783117
hs02571323888122
hs02571323989123
hs02571324093127

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
HhaIGCGCGCGCATTTATAGAAACCGGGGGCGCGG
ScrFICCNGGCCGGGATTTATAGAAACCGGGGGCGCGG
FnuDIICGCGCGCGATTTATAGAAACCGGGGGCGCGG
GlaIGCGCGCGCATTTATAGAAACCGGGGGCGCGG
PsuGIBBCGDCGCGGATTTATAGAAACCGGGGGCGCGG
FaiIYATRTATAATTTATAGAAACCGGGGGCGCGG
CauIICCSGGCCGGGATTTATAGAAACCGGGGGCGCGG
HpaIICCGGCCGGATTTATAGAAACCGGGGGCGCGG
SecICCNNGGCCGGGGATTTATAGAAACCGGGGGCGCGG

SNPs and Variation

Notes

× Warning! This gRNA contains polymeric runs 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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