Cat.No.：D26011A
Price: ￥
Specifications：
The Lba Cas12a (Cpf1) nuclease is derived from the ND2006 strain of the Myxococcus genus bacteria. It is a programmable RNA-mediated DNA endonuclease of the II type V CRISPR system and is a superior alternative editing tool for the mainstream Cas9 nuclease.
Compared with the traditional Cas9 protein, this Cas12a enzyme does not require an auxiliary tracrRNA. It can complete targeted recognition and cutting solely by relying on a 41–44 nt short-chain crRNA. This enzyme specifically recognizes the 5′-TTTV conserved PAM sequence in the genome and the cutting site is located 17–18 base pairs downstream of the PAM. After the cutting, a 5′ sticky protruding end is formed, which is more conducive to gene site-directed editing.
Both ends of the protein are fused with the SV40 nuclear localization signal (NLS). The dual NLS structure significantly improves the protein's nuclear entry efficiency and effectively enhances the success rate of gene editing both in vitro and in vivo, making it suitable for editing experiments on various biological samples.
1. Broader targeting range
Specifically recognizes TTTV with T-shaped PAM sequences, enabling precise targeting of genomic AT-rich regions that Cas9 cannot cover, significantly expanding the selection range of genome editing targets.
2. Simpler guide design
Only a single short-chain crRNA is needed for targeted guidance, without the need for tracrRNA, resulting in lower synthesis costs, simpler experimental construction processes, and high friendliness for beginners.
3. Excellent cutting performance
Unique sticky-end cutting mode, different from Cas9's blunt-end cutting, is more suitable for precise knockout, fragment insertion, and gene modification experiments with refined editing.
4. Efficient nuclear entry
The protein's N-terminal and C-terminal dual SV40 nuclear localization signals enhance nuclear transport efficiency in eukaryotic cells, effectively increasing overall editing activity.
5. Strong temperature adaptability
It has an ultra-wide active temperature range (16–48℃), maintaining high enzyme activity even at low temperatures, making it particularly suitable for live body gene editing experiments in cold-blooded animals such as zebrafish and African clawed frogs.
6. Dual enzyme activity versatility
It combines double-stranded DNA specific cis-cutting activity and single-stranded DNA non-specific trans-cutting activity, meeting the needs of gene editing while also being applicable to rapid nucleic acid detection in vitro and molecular diagnosis scenarios.
7. High purity and stability
Verified by SDS-PAGE electrophoresis, the protein purity is ≥90%. Each batch is strictly controlled for activity and purity, with strong protein stability and high experimental reproducibility, supporting high-demand scientific research experiments.
| Cat. No. | Component | Concentration | Specification | Quantity |
| B57011A | Lba Cas12a (Cpf1) | 1 μM | 70 pmol | 1 |
Buffer system: 300 mM NaCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol, pH=7.4 (25°C)
1. Long-term preservation: It is recommended to store at -25℃ to -15℃ under low-temperature refrigeration, which can maintain high enzyme activity for a long time;
2. Short-term use: It can be stored for up to 12 months at 4℃;
3. Precautions: Do not repeatedly freeze and thaw. It is recommended to pack and store the product as needed after receiving it to retain enzyme activity to the greatest extent; The freeze-dried powder version can be stored for a long time at 4℃ in a dry and sealed state.
This Lba Cas12a nucleases aptamer can be used in various CRISPR experimental systems and is a multi-functional research tool enzyme. Its core applications are as follows:
1. Precise gene knockout, gene editing and gene function verification in model organisms such as mammalian cells, zebrafish, and frog;
2. Specific site-directed enzymatic cleavage of double-stranded DNA in vitro, vector construction, and fragment modification experiments;
3. Detection of nucleic acids based on the trans-activity of Cas12a, diagnosis of pathogenic molecules, and development of high-throughput screening technologies;
4. High-throughput genomic library screening, target validation, and research on new editing systems in cutting-edge scientific scenarios.
In the in vitro cutting experiment, there were no cutting products or the cutting was incomplete. What could be the reasons for this?
Possible reasons include: Inappropriate molar ratio: It is recommended to maintain the molar ratio of Lba Cas12a : gRNA : target site at 10:10:1 or higher. Inappropriate design of gRNA sequence: It is necessary to verify whether the sequence and design of gRNA match the target site. Poor quality of gRNA: Verify the integrity of gRNA through gel electrophoresis. Inappropriate buffer system: Please use the recommended reaction buffer. Improper dilution method: For example, if an in vitro cleavage experiment using 100 µM high-concentration enzyme is conducted, it can be diluted to 1 µM with Lba Cas12a Diluent first before use; supplementing 10 mM DTT to the reaction system may help improve the cleavage efficiency.
How is the gRNA for Lba Cas12a designed and obtained?
The gRNA of Lba Cas12a is 41-44 nucleotides in length. A crRNA sequence complementary to the target site needs to be designed to ensure that there is a 5' TTTV PAM sequence on the opposite DNA strand of the target site. The gRNA can be obtained through in vitro transcription or chemical synthesis. It should be noted that the sgRNA synthesis kit for Spy Cas9 is not suitable for the synthesis of gRNA for Lba Cas12a.
What are the reaction conditions for Lba Cas12a?
It is recommended to use 1× NEBuffer™ r2.1 reaction buffer and incubate at 37℃ for the in vitro cleavage reaction. After the reaction is completed, the enzyme can be inactivated by incubation at 65℃ for 10 minutes.
Which species' genes can be edited by Lba Cas12a?
Lba Cas12a exhibits activity within the temperature range of 16–48℃. Due to its ability to maintain activity at lower temperatures, it is particularly suitable for gene editing in cold-blooded animals such as zebrafish and African clawed frogs. It can also be used for gene editing in conventional mammalian cells, and the dual NLS design enhances the nuclear entry efficiency.
For which experiments is the high concentration of Lba Cas12a (100 µM) suitable?
High-concentration liquid formulation (M0653T) is suitable for applications such as microinjection, electroporation and liposome transfection that require a relatively high enzyme concentration. When used in in vitro cleavage experiments, it can be diluted to 1 µM with Lba Cas12a Diluent before being used.
What are the uses of the trans-cutting activity of Lba Cas12a?
In addition to specifically cutting the double-stranded DNA of the target, Lba Cas12a also has the non-specific trans-activity of cutting single-stranded DNA. This characteristic can be applied to in vitro nucleic acid detection and diagnosis (such as CRISPR Diagnostics), generating detectable signals through the cleavage of the reporter probe.
