SpCas9 protein for precise editing
SpCas9 nuclease is a high-quality, recombinant protein that enables robust and reliable genome editing. This widely used CRISPR nuclease, complexed with a synthetic single guide RNA (sgRNA), can achieve high editing efficiencies. It features two nuclear localization signals (2NLS) to enhance its transport into the nucleus for precise gene editing.
Since its discovery in the Nobel Prize-winning publication, Cas9 nuclease has been used repeatedly in various applications spanning protein visualization studies, the generation of disease models, agriculture improvements, and most notably, in the cell and gene therapy space making it the gold standard in CRISPR gene editing. Of the different Cas9 nuclease variants, the most widely known is Streptococcus pyogenes (SpCas9). To maximize its efficacy, researchers and scientists know that complexing SpCas9 nuclease with synthetic sgRNA enables them to achieve desirably high editing efficiencies needed to be successful in their CRISPR experiments and therapeutic development. However, not all SpCas9 nuclease variants sourced from different companies provide the high efficiency needed to be successful at gene editing.
For over a decade, we have been a leader in providing CRISPR solutions that unlock new discoveries and treatments. As a leader, we provide a highly efficient SpCas9 nuclease that leads to the robust and reliable genome editing results you need. To maximize the efficacy of genome editing, our SpCas9 nuclease has two nuclear localization signals (2NLS) to enhance its transport from the cytoplasm into the nucleus. When complexed with our best-in-class Research sgRNA, SpCas9 nuclease precisely makes blunt end double-strand break roughly 3 bases nucleotides upstream of the protospacer adjacent motif (PAM) - in this case, SpCas9 nuclease PAM sequence is 5′-NGG-3′. The results speak for themselves, researchers and scientists have been using our SpCas9 nuclease and Research sgRNA to make groundbreaking discoveries and so can you.
Whether you are developing revolutionary therapies or conducting cutting-edge research, our SpCas9 nuclease provides the precision and reliability you need.
Our research SpCas9 nuclease has two nuclear localization signals (NLS) to boost transportation to the nuclease for gene editing. For additional product specifications, please see the following:
| Product Number | R20SPCAS9 |
| Concentration | 20 µM 10 mg/ml |
| Intended Use | This product is intended for research use only |
| Shipping | Cold Pack |
| Buffer Composition | 25-30 mM Tris-HCl*, 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4 |
| Source | E. coli |
| Purity | ≥ 95.0% |
Like other nucleases and RNA, SpCas9 and Research sgRNA should be used in an RNase-free and sterile environment. To avoid introducing contaminants, it is highly recommended that you use sterile filter pipette tips.
For designing SpCas9 guide RNA(s), use your favorite guide design platforms (e.g., Synthego's Design Tool, CHOPCHOP, Benchling, CRISPOR, or CRISPR RGEN Tools) and select SpCas9 nuclease, which has a 5'-NGG PAM sequence. After you design your SpCas9 guide RNA, use our easy-to-use order interface to order your guides. We will synthesize them resulting in high-quality synthetic guide RNA you complex with our SpCas9 nuclease to achieve your highest editing efficiency.
The following modifications are included on our SpCas9 sgRNA during synthesis:
2'-O-Methyl at 3 first and last bases, 3' phosphorothioate bonds between first 3 and last 2 bases
SpCas9 Nuclease |
hfCas12Max Nuclease |
||
|---|---|---|---|
| Description | The nuclease that led to Jennifer Doudna and Emmanuelle Charpentier's Nobel Prize. | Engineered from Cas12i to achieve higher on-target editing and minimized off-target editing. | |
| Size | 1368 amino acids | 1080 amino acids | |
| PAM Sequence (N = any nucleotide) | 5'-NGG-3’ PAM is 3’ of the target DNA sequence |
5'-TN-3' or 5'-TTN-3' | |
| DNA Cleavage | Blunt end cut 3nt upstream of PAM sequence | Staggered-cut: cleavage on the target strand occurs 24nt downstream from PAM, while the non-targeted strand is cut 14-16nt downstream. | |
| Endonuclease Domains | 97 - 103nt 20nt Target Sequence |
44 - 50nt 20nt Target Sequence |
|
| gRNA Components | crRNA + tracrRNA as a single guide RNA (sgRNA) | crRNA | |
| Variants or Origin | Streptococcus pyogenes (Wildtype) | Engineered from Cas12i | |
| CRISPR Enzyme Class | Type IIa CRISPR-Cas system of Parasutterella secunda Cas9 (PsCas9) | Type V CRISPR-Cas system of Cas12 | |
| Application | A wide range of applications spanning from research use to cell & gene therapy development for clinical trials. |
SpCas9 nuclease significantly impacts the cell and gene therapy space because it can rewrite the human genome. In 2023, the FDA approved the first SpCas9 nuclease CRISPR-based therapy, Casgevy, developed to treat sickle cell disease and beta-thalassemia. Explore other CRISPR-based therapies in clinical trials that are using SpCas9 nuclease.
Challenge: Subpar-sourced SpCas9 nucleases can cause trouble optimizing your CRISPR workflows, making it difficult to edit your target wasting valuable time and resources.
Solution: Our SpCas9 has demonstrated its ability to achieve consistently high editing efficiencies, enabling you to optimize your CRISPR workflows seamlessly and progress your work from early development to your essential downstream assay.
Challenge: Inefficient sourced SpCas9 cannot handle large, high-throughput screens to evaluate targets effectively for research and early therapeutic development.
Solution: Our SpCas9 nuclease can easily edit target loci on a high-throughput screen enabling you to identify hits to further evaluate in your important downstream assays.
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