| Description |
GenCRISPR™ ErCas12a Nuclease is an RNA-guided DNA endonuclease from Eubacterium rectale. 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 ErCas12a with a N-terminal NLS
expressed by E.coli
|
| Species |
Eubacterium rectale
|
| Tag |
C-terminal 8× His Tag
|
| Theoretical Molecular Weight |
~148 kDa
|
| Concentration |
4 mg/ml
|
| Storage Buffer |
20 mM Tris-HCl, 300 mM
NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol,
pH 7.4
|
| Storage & Stability |
Store
at -20 °C for up to 18 months from the date of manufacture. Avoid repeated
freeze-thaw cycles. Do not store below
-20 °C!
|
| Accession# |
WP_055225123.1
|
| 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 80 ng PCR product, 20 ng crRNA, and 40/60 ng GenCRISPR™ ErCas12a Nuclease (Cat. No. Z03762) for 60 min at 37°C results in a digestion efficiency of PCR product 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. Liu, Zhenyi, et al. "ErCas12a CRISPR-MAD7
for model generation in human cells, mice, and rats." The CRISPR journal 3.2 (2020): 97-108.Chen, Janice S., et al.
"CRISPR-Cas12a target binding unleashes indiscriminate single-stranded
DNase activity." Science
360.6387 (2018): 436-439.
3. Rojek, Johan, et al. "Mad7: An IP friendly
CRISPR enzyme." Authorea Preprints (2021). |
For laboratory research use only. Direct human use,
including taking orally and injection and clinical use are forbidden.
