Description
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.
|
Note
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).
2.
Gootenberg, Jonathan S., et al. "Nucleic acid detection
with CRISPR-Cas13a/C2c2." Science 356.6336 (2017):
438-442.
3.
Gootenberg, Jonathan S., et al. "Multiplexed and
portable nucleic acid detection platform with Cas13, Cas12a, and Csm6."
Science 360.6387 (2018): 439-444.
4.
Iwasaki, Roman S., and Robert T. Batey. "SPRINT: a
Cas13a-based platform for detection of small molecules." Nucleic
acids research 48.17 (2020): e101-e101. |
For laboratory research use only. Direct human use,
including taking orally and injection and clinical use are forbidden.
