LbCas12a: A Versatile CRISPR-Cas12a Enzyme for Precise Genome Editing
Unleash the Power of Precision Genome Editing with LbCas12a
LbCas12a, a CRISPR-Cas12a enzyme from Listeria borborealis, offers unparalleled precision and versatility for genome editing applications. Its unique features, including a PAM (protospacer adjacent motif) recognition sequence that is less stringent than that of Cas9, make it suitable for a wider range of target sites. Additionally, LbCas12a's robust RNase activity enables efficient collateral cleavage, expanding its utility for gene knockouts and large-scale genomic modifications.
Key Benefits of LbCas12a:
- High Fidelity: LbCas12a introduces minimal off-target edits, ensuring precise genomic modifications.
- Broad Target Specificity: LbCas12a recognizes a variety of PAM sequences, expanding its target range.
- Efficient RNase Activity: LbCas12a's robust RNase activity facilitates gene knockouts and large-scale genomic modifications.
Applications of LbCas12a:
- Gene Knockout and Knock-in: Precisely disrupt or introduce genes of interest.
- Point Mutations and Small Insertions/Deletions (InDels): Introduce specific mutations or modify gene sequences with ease.
- Large-Scale Genomic Modifications: Facilitate efficient genome-wide editing projects.
Empower Your Genome Editing Efforts with LbCas12a
LbCas12a's versatility and precision make it a valuable tool for a wide range of genome editing applications. Whether you're seeking targeted gene modifications or large-scale genomic alterations, LbCas12a delivers the precision and efficiency you demand.
|Source||Recombinant expression in Escherichia coli|
|Storage Conditions||-20 ±5°C|
|Molecular Weight||143.7 kDa|
|Storage Buffer||20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 20% glycerol, 1 mM DTT|
|Quality Control||No residual nucleases (exonucleases and endonucleases)|
|Shelf Life||12 months|
Cas12a, also recognized as Cpf1, stands out as a potent type II V CRISPR effector protein. Functioning as an RNA-guided endonuclease, it selectively binds to specific target DNA sites, executing precise cleavage under the guidance of single-stranded guide RNA. This versatile protein has found extensive application in the realm of gene editing, spanning microorganisms, plants, and animals, showcasing promising potential in the field of molecular diagnostics.
Cas12a's proficiency extends beyond its ability to recognize and cleave double-stranded DNA targets; it exhibits activated efficiency in cleaving arbitrary sequence single-stranded DNA (ssDNA). The strategic design of probes, adorned with fluorescence or other small molecule markers at both ends, facilitates signal amplification in DNA detection through CRISPR/Cas12a. Commonly employed detection methods involve real-time fluorescence and colloidal gold assays.
Noteworthy is the gRNA-specific recognition of the target region, rendering the CRISPR/Cas12a system a valuable "secondary screening" tool for scrutinizing DNA or DNA amplification products. This attribute contributes to the system's high specificity in molecular applications.
|No.||Component Name||Specifications (100 pmol)||Specifications (1000 pmol)||Specifications (2000 pmol)|
|1||10×Cas12 Buffer||250 µL||5 mL||5 mL|
|2||LbCas12a||10 µL (10 µM)||200 µL (10 µM)||100 µL (20 µM)|
Introduction to LbCas12a:
In the realm of genetic manipulation, LbCas12a emerges as a remarkable entity with a distinct set of capabilities. Operating under RNA mediation, LbCas12a adeptly recognizes the PAM sequence TTTN site. Its precision extends to the targeted cleavage, triggering a fascinating "collateral cleavage" phenomenon that slices through any sequence of single-stranded DNA. Harnessing the power of probe design embellished with fluorescent or other small molecule markers at both ends facilitates signal amplification, paving the way for sensitive DNA detection with a keen focus on the designated target. The inherent characteristics of this system contribute to its elevated sensitivity and robust specificity.
- In a comparison with its counterpart Cas9, the LbCas12a protein exhibits a smaller size, rendering it more amenable to efficient delivery into cells.
- The reaction temperature range spans a broad spectrum from 20 to 48°C.
- Post-target cleavage, LbCas12a showcases its unique "collateral cleavage" activity, capable of incising any single-stranded DNA sequence.
- Gene editing stands out as a primary application for LbCas12a, leveraging its precision and efficiency in genetic alterations.
- When coupled with isothermal amplification technology, this system facilitates rapid and effective nucleic acid detection.
The efficacy of LbCas12a is substantiated by SDS-PAGE electrophoresis, providing a tangible representation of its performance and molecular characteristics.