CRISPR/Cas12b service

Genome editing has always been a frontier and hot area of biological research. Genome editing tools based on CRISPR-Cas9 and CRISPR/Cas12a have been widely used in basic research and applications, such as gene function analysis, germplasm creation, gene therapy, etc. Compared with CRISPR-Cas9 and CRISPR/Cas12a systems, CRISPR/Cas12b is a universal and customizable system in plants, which can finally provide effective gene editing to achieve target gene activation and inhibition.

Figure 1. Protein domain structure of Cas12b. (Strecker et al., 2019)

As a dual-RNA-guided nuclease, Cas12b contains a single RuvC domain and requires both CRISPR RNA (crRNA) and transactivating crRNA (tracrRNAs), which can be combined as a single guide RNA (sgRNA), for DNA targeting. As a class 2 type V-B CRISPR system, Cas12b prefers T-rich protospacer adjacent motifs (PAMs) and generates staggered ends of DNA double-strand breaks. CRISPR/Cas12b provides a higher efficiency for gene activation and a wider target site for gene suppression in plant gene editing.

Figure 2. Schematic of Cas12b sgRNA design. (Strecker et al. 2019)

Advantages of the CRISPR/Cas12b System.

• Wide range of applications. Compared with the Cas9, Cas12b recognizes simpler PAM sequences, which can significantly increase the target range of the genome. In addition, it is reported that CRISPR/Cas12b system could realize multiple genome editing (Ming et al. 2020).
• High security. The Cas12b protein is smaller than Cas9 and Cas12a, so it is easier to achieve cell-to-cell delivery through viral vectors. Cas12b could hardly tolerate any mismatches between sgRNA and target DNA sequences, so it has less off-target effects than Cas-9 and Cas12a, which means it is safer for genome editing (Teng et al. 2018).
• High gene activation efficiency. In addition to retaining the inherent custom cutting and gene regulation capabilities of Cas12a, Cas12b can also improve the efficiency of gene activation (Ming et al. 2020).

Lifeasible is a world-class phytotechnology company with extensive experience in plant gene editing. We are devoted to optimizing the CRISPR system to achieve genetic modification. Lifeasible has developed a number of Cas12b vectors such as Alicyclobacillus acidoterrestris (Aac), Alicyclobacillus acidiphilus (Aa), Bacillus thermoamylovorans (Bth) and Bacillus hisashii (Bh) and plant transformation systems of tobacco, potato, cotton, pepper, barley, Arabidopsis, soybean, rice, which are readily available to accommodate the diverse needs of the clients.

CRISPR/Cas12b system experiment procedure

We hope that our technology and services can help you achieve new breakthroughs. For more information, please contact Lifeasible.