| Description |
GenCRISPR™ LbuCas13a Nuclease is an RNA-guided
RNA endonuclease from Leptotrichia
buccalis. 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 buccalis
|
| Tag |
Tag-free
|
| Apparent Molecular Weight |
~140 kDa, on SDS-PAGE under non-reducing
conditions
|
| Concentration |
4 mg/ml |
| Storage Buffer |
50 mM Tris-HCI, 600 mM NaCl, 0.1 mM TCEP, 50% glycerol, pH 7.5.
|
| 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# |
C7NBY4
|
| Appearance |
Clear, colorless liquid
|
| Purity |
≥ 90% 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
|
LbuCas13a 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.
5.
East-Seletsky,
Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable
guide-RNA processing and RNA detection." Nature 538.7624 (2016): 270-273. |
For laboratory research use only. Direct human use,
including taking orally and injection and clinical use are forbidden.
