20 Life Technologies
™
| Genome editing
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TECHNICAL PRODUCT BULLETIN GeneArt
®
CRISPR Nuclease Vector Kit
GeneArt
®
CRISPR Nuclease Vector Kit
Rapid and efficient genome editing from a single vector
Introduction
The Clustered Regularly Interspaced Short
Palindromic Repeats (CRISPR) and CRISPR-
associated (Cas) system is the latest addition
to the genome editing toolbox, offering a
simple, rapid, and efficient solution. The
CRISPR/Cas system is a prokaryotic adaptive
immune system that uses an RNA-guided
DNA nuclease to silence viral nucleic acids
[1]. The type II CRISPR/Cas system from the
bacterium Streptococcus pyogenes has been
modified to enable editing of mammalian
genomes [2, 3]. As a simple two-component
system composed of Cas9 protein and a non-
coding guide RNA (gRNA), the engineered
type II CRISPR/Cas system can be utilized to
cleave genomic DNA at a predefined target
sequence of interest.
The gRNA has two molecular components, a
target complementary CRISPR RNA (crRNA),
and an auxiliary trans-activating crRNA
(tracrRNA). The gRNA unit guides the Cas9
nuclease to a specific genomic locus and
the Cas9 protein induces a double-strand
break (DSB) at the specific genomic target
sequence (Figure 1). Following CRISPR/
Cas9-induced DNA cleavage, the DSB can
be repaired by the cellular repair machinery
using either non-homologous end joining
(NHEJ) or a homology-directed repair
mechanism. This product bulletin provides
detailed information about our GeneArt
®
CRISPR Nuclease Vector technology along
with recommendations for using the kits.
Target-specific crRNA
Target genomic loci
PAM
C G T A A A G C C A T A C G T A T A C T A C C
G C A T T T C G G T A T G C A T A T G A N G G
G G G G C C C A A
A
A U U U U
G
A
A
U
U U G A U
tracrRNA
A A A A A A
A
A
A
A
A
A
A
A
A
A
A
A
A A
A
G
G
G
G
G
G
G G G
G
G
G
G
G
G
G
G
G
G C C C C
C
C
C
C
C
C
C
C
U
U
U U U
U
U
U
U
U U U U
U
U U
U
U
U
A
A
A A
A
U
U
U
U
Cas9
Figure 1. A CRISPR/Cas9 targeted double-strand break. Cleavage occurs
on both strands, 3 bp upstream of the NGG proto-spacer adjacent motif (PAM)
sequence on the 3' end of the target sequence.
Find out more at lifetechnologies.com/crispr
PG1321-PJ5212-CO28372-CRISPR Technical Product Bulletin (Global).indd 1 11/15/13 1:23 PM
CRISPR-Cas9 Technical Resources
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