| Description |
GenCRISPR™ LbCas12a Nuclease is an
RNA-guided DNA endonuclease from Lachnospiraceae
bacterium. Cas12a (previously known as Cpf1) belongs to the Class 2 Type V CRISPR/Cas
system. Different from CRISPR/Cas9 system, the ribonucleoprotein (RNP) complex
of CRISPR/Cas12a system is formed by Cas12a and crRNA, it recognizes a T-rich protospacer
adjacent motif (PAM) and results in a staggered DNA double-strand break (DSB). After
the specific cleavage, Cas12a can also activate
collateral cleavage activity towards adjacent non-specific ssDNA sequences. Hence, Cas12a nuclease is a good alternative for Cas9 in certain
target DNA editing, and provides a novel strategy for DNA detection. |
crRNA-dependent DNA cleavage
|
| Source |
Recombinant Cas12a with a C-terminal NLS
expressed by E.coli
|
| Species |
Lachnospiraceae bacterium
|
| Tag |
C-terminal 6× His Tag
|
| Apparent Molecular Weight |
146 kDa, on SDS-PAGE under non-reducing
conditions
|
| Concentration |
4 mg/ml
|
| Storage Buffer |
10 mM Tris-HCl, 300 mM NaCl, 0.5 mM DTT, 50%
glycerol, pH 7.4.
|
| 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# |
6KL9_A
|
| Appearance |
Clear, colorless liquid
|
| Purity |
≥ 90% by SDS-PAGE
|
| Concentration by A280 |
4(± 10%) mg/ml
|
| Bioactivity (in vitro) |
≥ 90%
|
| Residual DNase |
Undetectable
|
| Residual RNase |
Undetectable
|
| Endotoxin Level |
< 0.1 EU/μg
|
Figure 1: In vitro digestion efficiency analysis by agarose gel electrophoresis. A 20 μl reaction in 1 × Cas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ LbCas12a Nuclease (Cat. No. Z03753) for 30 min at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.
| References |
1. Zetsche, Bernd, et al.
"Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas
system." Cell 163.3 (2015): 759-771.
2. Ledford, Heidi. "Alternative CRISPR system could improve
genome editing." Nature News
526.7571 (2015): 17.
3. Chen, Janice S., et al.
"CRISPR-Cas12a target binding unleashes indiscriminate single-stranded
DNase activity." Science
360.6387 (2018): 436-439. |
For laboratory research use only. Direct human use,
including taking orally and injection and clinical use are forbidden.
