High-fidelity Cas9 mRNA for therapeutic gene editing.
eSpOT-ON is an engineered high-fidelity Cas9 variant, designed for therapeutic gene editing. Delivered in mRNA format, eSpOT-ON provides superior on-target activity with low off-target effects. This nuclease has been validated in preclinical studies and is ready to support your next breakthrough in cell and gene therapy development.
eSpOT-ON mRNA is a next-generation high-fidelity Cas9 designed for precise and efficient genome editing in cell and gene therapy development. Derived from Parasutterella secunda Cas9 (PsCas9) and also known as engineered PsCas9 (ePsCas9).
This engineered mRNA delivers strong on-target activity while minimizing off-target effects, enabling safer and more reliable therapeutic editing. Producing staggered DNA cuts, eSpOT-ON mRNA enhances DNA repair and integration efficiency, supporting advanced CRISPR applications across multiple human cell types. Compatible with chemically modified sgRNA, eSpOT-ON mRNA helps accelerate workflows from early discovery to preclinical therapeutic development.
High-Fidelity Editing:
Strong on-target activity with minimal off-target effects for safer genome editing.
Optimized for mRNA Delivery:
Engineered for efficient translation in therapeutic applications, including LNP or viral vector systems.
Staggered DNA Cuts:
Improves DNA integration efficiency for targeted knock-ins and therapeutic gene correction.
Chemically Modified sgRNA Compatible:
Maximizes editing precision and efficiency when paired with optimized guide RNAs.
Validated Across Cell Types:
Demonstrates consistent performance in diverse human cell lines, including primary cells and iPSCs.
| Product Number | M20ESPOTON |
| Concentration | 1 µg/µl |
| Delivery Format | 1.5 ml plastic tube 5 x 1.5ml plastic tubes |
| Intended Use | This product is intended for research use only |
| Shipping | Cold Pack |
| Buffer Composition | 10 mM Tris-HCl (pH 7.5), 1 mM EDTA, RNase-free |
| Source | in vitro transcribed (IVT) |
| Purity | ≥ 80.0% RIN |
| Specification | eSpOT-ON Nuclease |
|---|---|
| Size | 1409 amino acids |
| PAM Sequence (N = any nucleotide) | 5'-NGG-3’ |
| DNA Cleavage | Staggered-cut. Cleavage on the target strand occurs 3 nt upstream of the PAM, while the non-target strand is cut 6-7 nt upstream of the PAM. |
| Endonuclease Domains | HNH and RuvC |
| gRNA Length | 109nt |
| Target Sequence Length | 22 nt |
| Enzyme Class | Type IIb CRISPR-Cas system of Parasutterella secunda Cas9 |
To design guide RNAs compatible with eSpOT-ON mRNA, you can use one of these guide design tools. If eSpOT-ON isn’t listed as a default option, simply input the 5’-NGG-3’ PAM and its corresponding staggered cut profile to design your eSpOT-ON gRNA. Once you’ve selected your target sequence, paste it into our ordering platform. Our team will incorporate the correct eSpOT-ON scaffold (constant region) during synthesis to deliver a complete, ready-to-use gRNA.
To support the integration of our high fidelity Cas9 mRNA into your CRISPR workflows, we offer the eSpOT-ON Controls Kit, a convenient way to evaluate performance before moving to your specific targets. The kit includes two validated human positive control sgRNAs and one non-targeting control, providing a reliable benchmark for testing the efficiency and specificity of this high-fidelity engineered Cas9.
The optimized full-length gRNA includes the following modifications:
2'-O-Methyl analog at the first 3 and last 3 bases and 3' phosphorothioate bonds between 3 first and last 4 bases
eSpOT-ON mRNA is an engineered high-fidelity Cas9 mRNA designed to deliver precise, reproducible genome editing with a strong safety profile. By minimizing off-target activity while maintaining efficient on-target performance, eSpOT-ON mRNA supports the development of CRISPR-based therapies aligned with clinical and regulatory expectations.
If you have additional questions please connect with a member of our team.
In contrast, eSpOT-ON protein is delivered as a pre-formed ribonucleoprotein (RNP) complex with its guide RNA, enabling immediate genome editing upon cellular entry and allowing precise control over editing conditions.
Both formats provide the same high-fidelity eSpOT-ON Cas9 activity. The optimal choice depends on your delivery strategy, editing timeline, and therapeutic or experimental requirements.
Legal Terms: eSpOT-ON Nuclease purchased from Synthego is only intended and licensed to be used solely with eSpOT-ON guide RNAs purchased from Synthego, and eSpOT-ON guide RNAs purchased from Synthego are only intended and licensed to be used solely with eSpOT-ON Nuclease purchased from Synthego. For more details, visit our patent page.
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