GenCRISPR™ LbCas12a Nuclease
GenCRISPR™ LbCas12a Nuclease

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.

GenCRISPR™ LbCas12a Nuclease

GenCRISPR™ LbCas12a Nuclease is an RNA-guided DNA endonuclease from Lachnospiraceae bacterium. Cas12-family enzymes exhibit two DNase activity modes; besides crRNA-guided cis-cleavage of cDNA targets, these nucleases also catalyze trans-cleavage of non-target ssDNA substrates introduced by target DNA, which has been exploited to develop sensitive technologies for nucleic acid detection.
Z03753
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Overview
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.

Applications
crRNA-dependent DNA cleavage

Properties
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

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

Examples
  • GenCRISPR™ LbCas12a Nuclease
    • GenCRISPR™ LbCas12a Nuclease

    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.


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. 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.