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GenCRISPR Cas13a (C2c2) Nuclease is an RNA-guided, RNA-targeting CRISPR enzyme, commonly referred to as C2c2. This enzyme is a member of Type VI CRISPR family containing a single protein effector with two HEPN domains. The Cas13a exhibits two RNase activities. The first activity, a cis- sequence-specific RNA guided cleavage, activates the secondary RNase activity of the enzyme, a non-specific trans- ribonuclease activity. This Cas13a protein can be applied to detect single RNA molecules with proven high specificity. Recently, Cas13a has been leveraged as a diagnostic nucleic acid detection tool. Using a combination of the Cas13a cis- and trans- RNase activity and a non-target RNA tagged with a cleavage reporter ((fluorescence), the Cas13 system provides a route for reporting of the presence of a specific single RNA sequence in a sample with high specificity. The GenScript Cas13a contains a His-tag at the C-terminal. |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
| Description |
GenCRISPR™ Cas13a (C2c2) Nuclease is an RNA-guided RNA endonuclease from Leptotrichia wadei. This recombinant nuclease is purified from E. coli expression and contains C-terminal His Tag. Cas13a (previously known as C2c2) belongs to the Class 2 Type VI CRISPR-Cas system and contains a single protein effector with two HEPN domains. Cas13a exhibits two distinct ribonuclease activities. Cas13a possesses sequence-specific RNA cleavage activity in vitro and in vivo. And after specific cleavage, Cas13a activates collateral cleavage activity towards adjacent non-specific ssRNA sequences. The two distinct ribonuclease activities of Cas13a enable guide-RNA processing and RNA detection. The RNA detection empowered with Cas13a-crRNA complex and quenched fluorescent ssRNA as reporter has been reported to be versatile, rapid and high-specific. |
|
crRNA-dependent RNA cleavage |
| Source |
Recombinant Cas13a expressed by E.coli |
| Species |
Leptotrichia wadei |
| Tag |
C-terminal His Tag |
| Apparent Molecular Weight |
~140 kDa, on SDS-PAGE under non-reducing conditions |
| Concentration | 4 mg/ml |
| Storage Buffer |
10 mM Tris-HCl, 300 mM NaCl, 1.0 mM DTT, 50% glycerol, pH 8.0. |
| Storage & Stability |
Store at -20 °C for up to 12 months from the date of manufacture. Avoid repeated freeze-thaw cycles. Do not store below -20 °C! |
| Accession# |
U2PWF1 |
| Appearance |
Clear, colorless liquid |
| Purity |
≥ 95% by SDS-PAGE |
| Activity |
≥ 85% by in vitro digestion of ssRNA |
| Concentration |
4 mg/ml ± 10% as analyzed by A280 |
| Residual DNase |
Undetectable |
| Residual RNase |
Undetectable |
| Endotoxin Level |
< 0.2 EU/μg |
LwaCas13a has sequence-specific RNA cleavage activity, and after specific cleavage, it activates collateral cleavage activity towards adjacent non-specific ssRNA sequences.
A 50 μL reaction containing ssRNA, Cas13a/crRNA complex, Fluorescent RNA reporter, and Cas13a Reaction Buffer is incubated for 30 minutes at 37 °C. After incubation, the fluorescence intensity is measured using a Microplate Reader (Excitation wavelength: 494 nm; Emission wavelength: 518 nm).
| References |
1. Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector." Science 353.6299 (2016). |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
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