描述
Award-winning GeneArt™ Platinum™ Cas9 Nuclease is the recombinant
Streptococcus pyogenes Cas9 (wt) protein purified from
E. coli that can be used for genome editing with CRISPR technology. Cas9 protein forms a very stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9 system. Incorporation of a nuclear localization signal (NLS) aids delivery to the nucleus, thus increasing the rate of genomic DNA cleavage.
With GeneArt Platinum Cas9 Nuclease you can:
• Achieve cleavage efficiencies of up to 85% in many cell lines (Liang, et al. 2015)
• Eliminate time consuming cloning steps
• Minimize off-target cleavage due to rapid clearance of the protein complex from the cell (Kim, et al. 2014)
• Streamline cell engineering by eliminating transcription and translation in the cell
Platinum Cas9 Nuclease is:
• Transfection ready, using either lipid-mediated transfection reagents or electroporation
• Manufactured under ISO 13485 Quality Standard
• Available in 2 concentrations:
--1 μg/μL for use in standard editing scenarios including cell lines such as HEK293 or HCT116 (Cat. Nos.
B25640 [25 μg] and
B25642 [10 μg])
--3 μg/μL for optimization of editing conditions in more difficult scenarios such as in primary or embryonic cell lines, or when screening multiple gRNA sequences at a time (Cat. No.
B25641)
--For custom sizes or concentrations, please
inquire.
How to obtain a gRNA sequence Use our
GeneArt CRISPR Search and Design Tool to search our database of >600,000 predesigned gRNA sequences specific to every gene in the human and mouse genomes. GeneArt predesigned gRNAs are optimized for gene knockout and typically target the first three transcribed exons per gene. Search results include recommendations based on minimizing potential off-target effects, potential binding sites, and exon maps with gRNA locations. This tool can also be used to analyze any sequence of interest to design unique CRISPR sequences.
How to make gRNA Once gRNA sequences have been selected, choose from three options for making gRNA:
1. The
GeneArt Precision gRNA Synthesis Kit for transfection-ready gRNA in as little as four hours including template assembly.
2. Order full-length, predesigned, transfection-ready
TrueGuide™ gRNAs.
3. Save time and effort and have our
GeneArt custom services team design, synthesize, and purify in vitro transcribed (IVT) gRNA sequences for you. To obtain a services quotation,
submit our online form.
Resources Demonstrated protocol: CRISPR-Cas9 microinjection in mice and zebrafish embryos References 1. Liang X, Potter J, Kumar S, Zou Y, Quintanilla R, Sridharan M, Carte J, Chen W, Roark N, Ranganathan S, Ravinder N, Chesnut JD. Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J Biotechnol. 2015; 208:44-53.
2. Kim S, Kim D, Cho SW, Kim J, Kim JS. Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res. 2014; 24(6):1012-9.
3. Liang X, Potter J, Kumar S, Ravinder N, Chesnut JD. Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA. J Biotechnol. 2017; 241:136-146.
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