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GenCRISPR™ Cas12a (Cpf1) Nuclease

The clustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonly present in archaea and bacteria. The CRISPR systems enable the host to specifically target and cleave foreign nucleic acids thus targeting infectiousviruses and plasmids. Recently, a type V CRISPR system has been identified in several bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an additional trans-activating crRNA (tracrRNA), and allow for targeting of additional genomic regions by cleaveing the target DNA proceeded by a short T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and purified. The nuclease contains nuclear localization sequence (NLS) at the C-terminus and 6× His-tag at the C-terminus.
$113.00
Z03502-50

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Overview
Description The clustered regularly interspaced short palindromic repeats, known as CRISPR systems are adaptive immune mechanisms commonly present in archaea and bacteria. The CRISPR systems enable the host to specifically target and cleave foreign nucleic acids thus targeting infectiousviruses and plasmids. Recently, a type V CRISPR system has been identified in several bacteria, the Cpf1 CRISPR from Prevotella and Francisella 1. In contrast to Cas9 systems, CRISPR/Cpf1 systems are smaller in size, do not require an additional trans-activating crRNA (tracrRNA), and allow for targeting of additional genomic regions by cleaveing the target DNA proceeded by a short T-rich protospacer-adjacent motif (PAM). On the other hand, the Cas9 system requires a G-rich PAM following the target DNA. Furthermore, Cas12a/Cpf1 introduces a staggered DNA double stranded break with a 4 or 5-nt 5’ overhang. Recombinant Acidaminococcus sp. BV3L6 Cas12a (cpf1) nuclease is expressed in E. coli and purified. The nuclease contains nuclear localization sequence (NLS) at the C-terminus and 6× His-tag at the C-terminus.
Key Features High knockout efficiencies: Consistent high performance in in-vitro plasmid cleavage test.
Time-saving: no need for transcription and translation. 
DNA-free: no external DNA added to system.
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Applications
crRNA -dependent double-stranded DNA cleavage.
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Properties
Source Recombinant Cas12a with an C-terminal NLS expressed by E.coli.
Species Acidaminococcus sp. (strain BV3L6)
PAM TTTV
Tag C-terminal 6× His Tag
Apparent Molecular Weight ~150 kDa, on SDS-PAGE under reducing conditions.
Concentration 4(±10%) mg/ml
Active temperature This Cas12a is active at 37℃.
Storage Buffer 20 mM Sodium Acetate, 500 mM NaCl, 1 mM TCEP，0.1 mM EDTA, 50% glycerol, PH6.0.
Storage & Stability This product remains stable for 18 months at -20℃. Avoid repeated freeze-thaw cycles.
Accession# U2UMQ6
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Quality Control Specifications
Appearance Clear, colorless liquid
Purity ≥ 90% by SDS-PAGE
Bioactivity ≥ 90%
Concentration 4(±10%) mg/mL
Residual DNase Undetectable
Residual RNase Undetectable
Endotoxin Level < 0.1 EU/μg of the protein by gel clotting method
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GenCRISPR™ Cas12a (Cpf1) Nuclease

A 20 μl reaction in 1xCas12a Nuclease Reaction Buffer containing 60 ng linearized plasmid, 10 ng crRNA, and 100 ng GenCRISPR™ Cas12a (Cpf1) Nuclease for 30 mins at 37°C results in a digestion efficiency of linearized plasmid 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.    Ledford, Heidi. "Alternative CRISPR system could improve genome editing." Nature News 526.7571 (2015): 17. 
3.    “Cpf1 Takes CRISPR Bigger by GoingSmaller.”

--> 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.    “Cpf1 Takes CRISPR Bigger by GoingSmaller.”
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For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.