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CRISPR-Cas9 Technology Information
Revolutionizing the field of genome editing
The transformative CRISPR-Cas9 technology is revolutionizing the field of genome editing. Able to achieve highly flexible and specific targeting, the CRISPR-Cas9 system can be modified and redirected to become a powerful tool for genome editing in broad applications such as stem cell engineering, gene therapy, tissue and animal disease models, and engineering disease-resistant transgenic plants. We've put together a collection of resources that we hope will give you the confidence to get started and to continuously improve your research.
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. Derived from components of a simple bacterial immune system, the CRISPR-Cas9 system permits targeted gene cleavage and gene editing in a variety of eukaryotic cells, and because the endonuclease cleavage specificity in CRISPR-Cas9 system is guided by RNA sequences, editing can be directed to virtually any genomic locus by engineering the guide RNA sequence and delivering it along with the Cas endonuclease to your target cell. The CRISPR-Cas9 system has great promise in broad applications such as stem cell engineering, gene therapy, tissue and animal disease models, and engineering disease-resistant transgenic plants.
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).
In bacteria CRISPR loci are composed of a series of repeats separated by segments of exogenous DNA (of ~30 bp in length), called spacers. The repeat-spacer array is transcribed as a long precursor and processed within repeat sequences to generate small crRNAs that specify the target sequences (also known as protospacers) cleaved by Cas9 protein, the nuclease component of CRISPR system. CRISPR spacers are then used to recognize and silence exogenous genetic elements at the DNA level. Essential for cleavage is a three-nucleotide sequence motif (NGG) immediately downstream on the 3’ end of the target region, known as the protospacer-adjacent motif (PAM). The PAM is present in the target DNA, but not the crRNA that targets it (Figure 1).
Upon binding to the target sequence, the Cas9 protein induces a specific double-strand break. Following DNA cleavage, the break is repaired by cellular repair machinery through non-homologous end joining (NHEJ) or homology-directed repair (HDR) mechanisms. With target specificity defined by a very short RNA-coding region, the CRISPR-Cas9 system greatly simplifies genome editing.
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.
Available GeneArt CRISPR-Cas9 genome-editing tools
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.
Currently we are the only company to offer the complete suite of genome editing reagents. We offer our state-of-the-art online Invitrogen™ CRISPR Search and Design Tool along with Invitrogen™ CRISPR-Cas9 editing products in four formats: CRISPR-Cas9 protein, CRISPR-Cas9 mRNA, CRISPR-Cas9 plasmid, and CRISPR library services. These gene-editing solutions are paired with optimal cell culture reagents, delivery methods, and analysis tools, based on your application and cell type.
Choose the optimized and validated toolset that’s right for you, and bypass the trial and error.
|Format||CRISPR Protein||CRISPR mRNA||CRISPR plasmid||CRISPR Libraries|
|Product name||GeneArt™ Platinum™ Cas9 Nuclease||GeneArt™ CRISPR Nuclease mRNA||GeneArt™ CRISPR Nuclease Vector||GeneArt™ CRISPR Arrayed Libraries|
|gRNA design||Use the GeneArt CRISPR Search and Design Tool for optimal design and minimal off-target effects. Go to thermofisher.com/crisprdesign|
|gRNA synthesis||GeneArt Precision gRNA Synthesis Kit||GeneArt Precision gRNA Synthesis Kit||DNA oligo cloned into plasmid||n/a|
|Reporter-based enrichment||Sold separately GeneArt™ Genomic Cleavage Selection Kit||Sold separately GeneArt™ Genomic Cleavage Selection Kit||✓ included; all-in-one expression plasmid||n/a|
|No promoter constraint||✓||✓||CMV promoter|
|Ready-to-use||✓||✓||Required cloning step||Ready-to-use lentiviral particles|
|No random integration concern||✓||✓||Concern||Stable expression of CRISPR-Cas9 system|
|Microinjection ready||✓||✓||Larger payload size|
|Multiplexing & screening capable||✓||✓||Larger payload size||High-throughput screening|
|Ready-to-act, stable RNP complex||✓|
Knockout and knock-in
|Knockout and knock-in||Knockout and knock-in||Loss of function screening|
|Delivery method*||Lipofectamine CRISPRMAX||Lipofectamine MessengerMAX||Lipofectamine 3000|
|*For the most efficient transfection of primary cells, stem cells, and difficult-to-transfect cells, use the Invitrogen Neon Transfection System.|
Not sure which CRISPR-Cas9 format to use? Let us help you ›
In this free webinar series you will learn tips and tricks for CRISPR delivery, how to best isolate edited clones, and downstream detection and validation methods for successful CRISPR gene editing.
The ultimate guide for getting started with CRISPR.
Intended as an introduction to genome editing, the resource guide covers the technologies available today for gene editing, along with methods for design, delivery and detection of edited cells.
CRISPR gRNA Design Tool
Our new simple, CRISPR Design tool provides access to >600,000 predesigned CRISPR gRNAs, options for de novo gRNA design, and recommendations based on potential off-target effects for each CRISPR sequence.
Demonstrated protocol: Engineering stem cells with CRISPR-Cas9
Using the right tools, editing stem cells can be done quickly and efficiently. Our free protocol offers a streamlined, robust, step-by-step guide for engineering both gene knock-in and knockout in induced pluripotent stem cells.
Demonstrated protocol: CRISPR-Cas9 microinjection in mice and zebrafish embryos
Strategies and tools for optimal editing efficiencies in mice and zebrafish. This step-by-step protocol was designed to guide you through efficient CRISPR embryo microinjections in mice and zebrafish.
Read the what, why, and how of CRISPR genome editing and its use in myriad endeavors including cell and protein analysis.
As genome-editing research increases in popularity, it is important to identify fast and cost-effective screening techniques and efficient editing protocols. Liang et al, describe methods for rapid synthesis of the sgRNA and for delivery of the Cas9 protein/gRNA ribonucleoprotein complexes into various mammalian cells either through electroporation or liposome-mediated transfection.
See if YOU can sort out fact from fiction and best this gene editing brainteaser.
Flyer: CRISPR Total Solution
We’re continually expanding our suite of genome editing products to span the entire cell engineering workflow, from reagents for cell culture, transfection, and sample preparation to kits for genome modification and detection and analysis of known genetic variants. These gene editing solutions are paired with optimal cell culture reagents, delivery methods, and analysis tools, based on your application and cell type.
It’s your time to change the world. And it’s our promise to empower you with cell engineering tools and solutions that help you get results even faster. Increasing your chances of success at every step.
The world’s largest collection of CRISPR engineered cell lines, great for rapid hypothesis testing. Powerful CRISPR models enabling clear results to drive your research forward.
For Research Use Only. Not for use in diagnostic procedures.