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GenCRISPR™ ErCas12a Nuclease

GenCRISPR™ ErCas12a Nuclease is an RNA-guided DNA endonuclease from Eubacterium rectale. 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.
$189.00
Z03762-100

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Overview
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.
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Applications
crRNA-dependent DNA cleavage
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Properties
Source Recombinant ErCas12a with a N-terminal NLS expressed by E.coli
Species Eubacterium rectale
PAM YTTV
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 18 months from the date of manufacture. Avoid repeated freeze-thaw cycles.
Accession# WP_055225123.1
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Quality Control Specifications
Appearance Clear, colorless liquid
Purity ≥ 90% (by SDS-PAGE)
Concentration 4 (± 10%) mg/ml
Bioactivity ≥ 90%
Residual DNase Undetectable
Residual RNase Undetectable
Endotoxin Level < 0.1 EU/μg
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GenCRISPR™ ErCas12a Nuclease

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. »

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Background
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).
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For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.