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E-GEOD-57650 - RNA-guided Location and Function of the Cas9 CRISPR Endonuclease in Mammalian Cells

Status
Released on 29 August 2014, last updated on 10 December 2014
Organism
Mus musculus
Samples (8)
Protocols (7)
Description
The clustered regularly interspaced short palindromic repeat (CRISPR)-associated enzyme Cas9 is an RNA-guided nuclease that has been widely adapted for genome editing in eukaryotic cells. However, the in vivo target specificity of Cas9 is poorly understood and most studies rely on in silico predictions to define the potential off-target editing spectrum. Using chromatin immunoprecipitation followed by sequencing (ChIP-seq), we delineate the genome-wide binding panorama of catalytically inactive Cas9 directed by two different single guide (sg) RNAs targeting the Trp53 locus. Cas9:sgRNA complexes are able to load onto multiple sites with short seed regions adjacent to 5’NGG3’ protospacer adjacent motifs (PAM). Examination of dmCas9 binding sites using two Trp53 targeting sgRNAs in Arf -/- MEF cell line (mouse).
Experiment type
ChIP-seq 
Contacts
Abba Malina, Hisashi Miura, Jerry Pelletier, Regina Cencic, Robert Francis
Citation
Protospacer adjacent motif (PAM)-distal sequences engage CRISPR Cas9 DNA target cleavage. Cencic R, Miura H, Malina A, Robert F, Ethier S, Schmeing TM, Dostie J, Pelletier J. , PMID:25275497
MINSEQE
Exp. designProtocolsVariablesProcessedSeq. reads
Files
Investigation descriptionE-GEOD-57650.idf.txt
Sample and data relationshipE-GEOD-57650.sdrf.txt
Processed data (2)E-GEOD-57650.processed.1.zip, E-GEOD-57650.processed.2.zip
Links