22 Life Technologies
™
| Genome editing
GeneArt
®
CRISPR Nuclease mRNA System
High-efficiency, multiplex-compatible genome
editing tool for a broad range of cell types
PRODUCT BULLETIN GeneArt
®
CRISPR Nuclease mRNA System
Introduction
Clustered regularly interspaced short palindromic
repeats (CRISPRs) and CRISPR-associated (Cas) proteins
have revolutionized the field of cell engineering. Derived
from components of a simple bacterial immune system,
the CRISPR-Cas9 system can be manipulated and
redirected for highly flexible but specific genome editing
in eukaryotes. The CRISPR-Cas9 system is composed
of a short noncoding guide RNA (gRNA) that has two
molecular components: a target-specific CRISPR
RNA (crRNA) and an auxiliary trans-activating crRNA
(tracrRNA). The gRNA unit guides the Cas9 protein to
a specific genomic locus via base pairing between the
crRNA sequence and the target sequence (Figure 1).
Upon binding to the target sequence, the Cas9 protein
induces a specific double-strand break. Following
CRISPR-Cas9–induced DNA cleavage, the break can be
repaired by the cellular repair machinery using either
non-homologous end joining (NHEJ) or a homology-
directed repair mechanism. With target specificity
defined by a very short RNA-coding region, the
CRISPR-Cas9 system greatly simplifies genome editing
and has great promise in broad applications such as
stem cell engineering, gene therapy, tissue and animal
disease models, and engineering disease-resistant
transgenic plants.
In the GeneArt
®
CRISPR Nuclease mRNA System,
crRNA and tracrRNA are expressed together as a
single gRNA that mimics the natural crRNA-tracrRNA
hybrid in bacterial systems. The gRNA is encoded by a
custom-synthesized DNA fragment containing either a
U6 or T7 promoter. GeneArt
®
CRISPR U6 Strings
™
DNA
is synthesized with a U6 promoter and can be directly
introduced into mammalian cells for expression of the
gRNA. GeneArt
®
CRISPR T7 Strings
™
DNA is synthesized
with a T7 promoter and is used as a template for in vitro
transcription of the gRNA. This target-specific gRNA is
cotransfected into cells with GeneArt
®
CRISPR Nuclease
mRNA, which encodes the Cas9 endonuclease. The
Target genomic locus
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.
Further your knowledge by downloading the latest technical product guide or
viewing one of our on-demand webinars at your leisure.
Did you know?
You can find a lot of data and application examples in our
GeneArt
®
CRISPR Nuclease mRNA product bulletin.
CRISPR-Cas9 technical resources
Let our knowledge work for you
