LbCas12a Nuclease

Product Introduction

LbCas12a (formerly known as LbCpf1) is a Type V-A CRISPR nuclease derived from the bacterium Lachnospiraceae bacterium ND2006, representing a class of gene editing tools distinct from the Cas9 system. Comprising approximately 1228 amino acids, this protein possesses a unique domain organization and biochemical characteristics, demonstrating application value complementary to SpCas9 in the field of genome editing.

The most notable features of LbCas12a lie in its working mechanism: First, it relies solely on crRNA guidance to recognize target sequences, without the need for tracrRNA assistance, simplifying the design of RNA components. Second, it recognizes the T-rich PAM sequence 5'-TTTV-3' (where V represents A, C, or G). This characteristic enables it to effectively target AT-rich genomic regions that are difficult for SpCas9 to cover, expanding the editable range across the entire genome. Third, LbCas12a generates 5-7 nt 5' overhangs (sticky ends) after cleaving DNA, rather than the blunt ends produced by Cas9, a feature that may facilitate directional gene insertion. Additionally, LbCas12a possesses trans-cleavage activity: after specifically recognizing and cleaving the target DNA, its nuclease domain is activated and can non-specifically cleave any single-stranded DNA molecules present in the system.

Product Specifications

Product Specifications

Application Scenarios

Gene Editing in AT-Rich Regions: Targeting T-rich PAM sites.

Precise Gene Knock-in: Sticky ends facilitating directional DNA fragment insertion.

Multiplex Gene Editing: Simultaneous editing of multiple targets using a single crRNA array.

Nucleic Acid Detection (DETECTR): Utilizing trans-cleavage activity for pathogen detection.

Plant Genome Editing: Suitable for plant genomes with high AT content.

Gene Therapy Research: Alternative to SpCas9 for specific target sites.

References

Zetsche B, et al. (2015). Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell. 163(3):759-771.

Chen JS, et al. (2018). CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 360(6387):436-439.

Strohkendl I, et al. (2018). Kinetic basis for DNA target specificity of CRISPR-Cas12a. Mol Cell. 71(5):816-824.