Description |
GenCRISPR™ ErCas12a Nuclease is an RNA-guided DNA endonuclease fromEubacterium 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.
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crRNA-dependent DNA cleavage
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Source |
Recombinant ErCas12a with a N-terminal NLS expressed byE.coli
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Species |
Eubacterium rectale
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Tag |
C-terminal 8× His Tag
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Theoretical Molecular Weight |
~148 kDa
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Concentration |
4 mg/ml
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Storage Buffer |
20 mM Tris-HCl, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol, pH 7.4
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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!
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Accession# |
WP_055225123.1
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Appearance |
Clear, colorless liquid
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Purity |
≥ 90% (by SDS-PAGE)
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Concentration by A280 |
4 (± 10%) mg/ml
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Bioactivity (in vitro) |
≥ 90%
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Residual DNase |
Undetectable
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Residual RNase |
Undetectable
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Endotoxin Level |
< 0.1 EU/μg
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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."Cell163.3 (2015): 759-771. 2. Liu, Zhenyi, et al. "ErCas12a CRISPR-MAD7 for model generation in human cells, mice, and rats."The CRISPR journal3.2 (2020): 97-108.Chen, Janice S., et al. "CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity."Science360.6387 (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.