Sequences  of A, B and C on- and off-target genomic sites from Hek293T cells treated with nickase Cas9 and target sgRNAs
Homo sapiens
Sequencing from Guilinger, Thompson & Liu. Nature Biotechnology 2014.  Three new target sites (sites A, B, and C) were chosen in the human genome that each had potential genomic off-target sites containing one identical or nearly identical half-site.  The three on-target sites and their highly similar off-target sites were amplified and sequenced from Hek293 cells treated with a mock transfection of a Cas9 nickase expression plasmid and either two sgRNAs spaced 15 bp apart targeting genomic site A, two sgRNA spaced 24 bp apart targeting genomic site B, or two sgRNA spaced 23 bp apart targeting genomic site C.  All genomic target sites (for A, B and C) were combined together for this one type of Cas9 treatment.
sex
female
tissue
Hek293
isolate
Hek293
age
?
biomaterial_provider
GenTarget
Cas9 expression vector
Cas9 nickase
gRNA targets
A, B and C
Cas9 expression vector (ng)
700
gRNAs expression vector (ng)
250
genomic sites amplifed
A, B and C
on-target sites
SiteA_On;SiteB_On;SiteC_On
off-target sites
SiteA _Off1-3;SiteB _Off1-3;SiteC _Off1-4
BioSampleModel
Human