Since its debut in 2012, CRISPR technology has achieved groundbreaking advancements in diverse fields such as gene functional studies, drug target screening, genetic disease therapy, cancer research, and crop breeding. Its mechanism of action is as follows: Cas proteins (e.g., the widely used Cas9) are guided by a guide RNA (gRNA) to precisely locate specific DNA sequences. Upon reaching the target site, Cas9 binds to the DNA and induces a double-strand break (DSB). In response to this damage, cells activate intrinsic DNA repair pathways, primarily non-homologous end joining (NHEJ) or homology-directed repair (HDR). By leveraging these endogenous repair mechanisms, scientists can introduce desired sequence modifications at targeted genomic loci, enabling precise genome editing.
Currently, CRISPR technology enables unprecedented precision in modifying DNA, including the insertion, deletion, or replacement of specific gene segments. This advancement not only facilitates more convenient and accurate gene correction but also opens new avenues for gene therapy, agricultural biotechnology, and synthetic biology.
Figure 1. YEASEN Gene Editing Solution
To support the aforementioned gene editing workflow, YEASEN offers high-quality products such as Cas proteins, along with customized services for protein engineering and GMP-scale production. These products span the entire process from experimental design to implementation, aiming to assist researchers in conducting gene editing experiments with greater efficiency and accuracy. Below, we highlight several key products.
sgRNA Synthesis Kit：Hifair® Precision sgRNA Synthesis Kit (Cat#11355ES)
This kit enables a highly streamlined application process. Users need only design a specific upstream primer and combine it with the reagents provided in the kit to synthesize 20–100 μg of high-quality sgRNA within 4 hours. The resulting sgRNA exhibits high yield, high purity, and high activity, significantly enhancing editing efficiency and reducing off-target effects, thereby ensuring high-efficiency and high-accuracy gene editing.
【Performance Data】
- High Yield: Single-tube reaction yields 20–100 μg of sgRNA within 4 hours.
Figure 2. sgRNA synthesis yield at different time points
- Broad Applicability: High yields achieved across various sgRNA sequences.
Figure 3. sgRNA synthesis yield with different sequences
- High Purity: Electropherogram (Agilent 2100) shows a single, clean sgRNA band.
Figure 4. Purity of synthesized sgRNA (analyzed by Agilent 2100)
- High Activity: The synthesized sgRNA effectively guides Cas9 to cleave target DNA.
Figure 5. sgRNA cleavage efficiency
Cas9 Protein: Cas9 Nuclease (Cat#14701ES)
This Cas9 nuclease is derived from Streptococcus pyogenes and is an RNA-dependent endonuclease capable of specifically cleaving double-stranded DNA . Cas9 cleavage occurs within the target sequence, three nucleotides upstream of the PAM (NGG) site. The protein has been codon-optimized and engineered with nuclear localization signals (NLS), and is recombinantly expressed in E. coli. It demonstrates high editing efficiency and is suitable for gene modification in various cell types (e.g., hematopoietic stem cells, T cells), as well as for molecular diagnostics, such as pathogen detection.
【Performance Data】
A：In vitro cleavage assay: Three batches showed cleavage activity exceeding 98%.
B：In vivo gene knockout: Knockout efficiency comparable to that of a leading international brand.
Figure 6. In vitro cleavage activity and gene knockout efficiency of Cas9 Nuclease
Cas12a Protein: ArCas12a Nuclease (Cat#14702ES)
ArCas12a nuclease, derived from the CRISPR system of Agathobacter rectalis (formerly known as Cpf1), is a single-protein enzyme comprising 1263 amino acids. Lacking the HNH domain, ArCas12a utilizes its RuvC domain alone to recognize a 5’-T-rich PAM under the guidance of CRISPR RNA (crRNA), initiating cleavage of target DNA. It has been successfully applied in genome editing of numerous mammalian and plant species. Additionally, ArCas12a exhibits trans-cleavage (collateral cleavage) activity, enabling non-specific cleavage of ssDNA in the reaction system. Compared to LbCas12a or AsCas12a, ArCas12a demonstrates superior thermostability (25–55°C), making it suitable for both genome editing and nucleic acid detection applications.
【Performance Data】
- Efficient in vitro dsDNA cleavage
Figure 7. Cis-cleavage activity of ArCas12a. M：Marker；C：Template dsDNA.
[Note]: Under crRNA guidance, ArCas12a efficiently cleaves a 600 bp dsDNA substrate into two fragments (200 bp and 400 bp).
- Strong trans-cleavage activity, suitable for nucleic acid detection
Figure 8. Trans-cleavage activity of ArCas12a
Note: Using dsDNA as the target, ArCas12a, crRNA (with a spacer complementary to the template), and a ssDNA reporter probe (fluorescently labeled) are introduced. Upon formation of the ArCas12a–crRNA–target DNA complex, trans-cleavage activity is activated, cleaving the ssDNA reporter and generating a fluorescent signal. Results confirm that the ArCas12a complex exhibits robust trans-cleavage activity.
Cas12b Protein: AapCas12b Nuclease (Cat#14808ES)
AapCas12b nuclease (also known as C2c1) is a tracrRNA:crRNA- or sgRNA-guided DNA endonuclease derived from Alicyclobacillus acidophilus. In the presence of a PAM sequence (TTN) on the target dsDNA, AapCas12b specifically cleaves the dsDNA, generating sticky ends. It can also cleave ssDNA targets in a PAM-independent manner. Both dsDNA and ssDNA targets can activate the trans-cleavage (collateral cleavage) activity of AapCas12b. Upon formation of the ternary complex (AapCas12b–sgRNA–target DNA), the enzyme gains non-specific ssDNA cleavage activity, degrading any ssDNA in the system. The optimal cleavage temperature for AapCas12b is 60°C, exhibiting higher thermostability than AacCas12b, making it ideal for integration with isothermal amplification techniques such as LAMP in developing CRISPR-based diagnostic assays.
【Performance Data】
- Cis-cleavage activity: Performance comparable to leading international brand
Figure 9. Validation of cis-cleavage activity of AapCas12b Nuclease
Note: In a 20 μL reaction containing dsDNA target with PAM, sgRNA, Cas12b, and 1× reaction buffer, incubated at 60°C for 30 min, followed by inactivation at 85°C for 5 min. Agarose gel electrophoresis confirmed efficient cleavage by three batches of AapCas12b, with performance matching that of brand T*.
- Trans-cleavage activity: Efficient cleavage of ssDNA in solution
Figure 10. Validation of trans-cleavage activity of AapCas12b Nuclease
Note: In a 20 μL reaction containing dsDNA target, sgRNA, Cas12b, fluorescent ssDNA reporter probe, and 1× reaction buffer, incubated at 60°C for 1 hour with fluorescence measured every 30 seconds. Fluorescence signal release was observed only when all components (target, sgRNA, Cas12b) were present, confirming trans-cleavage activity.
CRISPR gene editing technology is currently undergoing rapid development, with the field continuously maturing and evolving. As scientific research and technological applications advance, it is anticipated that more innovative gene editing tools will emerge, further expanding the scope and potential of this transformative technology.
For any inquiries or interests in gene editing applications—whether in basic research, drug development, agricultural biotechnology, or synthetic biology—please do not hesitate to contact us. We are committed to providing cutting-edge technical support and services to explore the vast possibilities offered by gene editing.
Ordering Information
|
Application |
Product Type |
Product Name |
Catalog Number |
|
General-purpose |
SpCas9 with NLS |
14701ES |
|
|
Compact size for delivery |
Cas12a |
14702ES |
|
|
Cas12b |
14808ES |
||
|
sgRNA preparation |
sgRNA synthesis |
11355ES |
|
|
sgRNA purification |
12602ES |
Extended Reading
Unraveling the Mechanism of Gene Editing: A Primer on CRISPR Technology
CRISPR/Cas Systems: Biological Formats and Delivery Strategies for Genome Editing