Through the years, many promising tools for genome editing have been developed including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR-associated protein 9 (Cas9). ZFN and TALENs target genome modification methods, design is costly and time-consuming, limiting their widespread use, especially for large-scale, high-throughput studies. CRISPR-Cas9 is a unique technology that enables geneticists and medical researchers to study, alter, create, and recreate highly complex pathways, DNA sequences, genes, and natural biological systems. It is currently the simplest, most versatile, and precise method of genetic manipulation and is therefore causing a buzz in the science world3. It has been widely used in the study of genome editing in broad applications such as stem cell engineering, gene therapy, tissue and animal disease models, and engineering disease-resistant transgenic plants, which has greatly improved the understanding of tumor genomics in people.
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ACROBiosystems focuses on the field of cell and gene therapy. As a leading supplier of recombinant proteins, ACROBiosystems has launched the CAS series nuclease including Cas9 and Cas12a. These proteins are mainly used for targeted gene editing to provide high editing efficiency.
The high enzyme activity is verified by in vivo/in-vitro experiments: In-vitro fragment cleavage efficiency >90%, which is facilitating genome editing with CRISPR technology.
Available in high concentrations of CAS-9 nuclease:10mg/ml for optimization of editing conditions in more difficult scenarios
High purity:SDS-PAGE & SEC-MALS verified purity > 90%.
Incorporation of nuclear localization signals (NLS) aids delivery to the nucleus, increasing the rate of genomic DNA cleavage
No residual RNase: The production process is strictly free from RNase pollution
Enzyme activity and purity have been inspected in batches. High stability and consistency between batches.
The interactions between prokaryotes and the viruses that infect them have evolved, leading to a wide diversity of CRISPR-Cas systems. CRISPR-Cas systems are generally divided into two categories (Class 1 and Class 2). To date, most researchers have used the Type 2 CRISPR-Cas system, and in this class, the most studied type II is the CRISPR-Cas9 system.
CRISPR-Cas9 related applications
- Applications in CAR-T therapy:
1) Generation of universal allogeneic CAR-T cells; 2) Enhancement of CAR-T cell function
- Application of TCR-T cell therapy:
α and β endogenous TCR Gene replacement with artificial tumor-specific TCR sequences for genes
- suppression of immune checkpoint signaling pathways
- screening of new targets for tumor immunotherapy
How does CRISPER-Cas9work
CRISPR/Cas9 system involves two essential components: target-specific CRISPR gRNA and Cas9 nuclease. In eukaryotic systems, CRISPR/Cas9 is used for genomic editing through specific targeting of DNA by sgRNA, a combination of the CRISPR RNA (crRNA) and the trans-activating crRNA (tracrRNA), mediated through base pairing over the ~20-nt guide sequence . Cas9 recognizes a very short conserved sequence (a few nucleotides in length) adjacent to the guide sequence called the “protospacer adjacent motif” (PAM). Once directed to the DNA target site, Cas9 generates a double-strand break (DSB) that can be repaired either through the indel mutation-introducing non-homologous end-joining (NHEJ) or the high-fidelity homologous directed repair (HDR), resulting in gene knockout effects or template-dependent gene replacement.
CRISPR-Cas9 regulatory mechanism: Repair of double-stranded DNA breaks by non-homologous end ligation (NHEJ) or homologous directed repair (HDR) endogenously
The high enzyme activity is verified by in vivo/in-vitro experiments
NLS-Cas12a Nuclease (MALS verified)（Cat. No.CAA-L5149
）substrate cutting efficiency>90%,the cleavage activity is comparable to that of competing products
Measured by its ability to cleave a targeted DNA substrate. Cas12a achieves >90% substrate cleavage, comparable to competing products
The enzyme activity of GENPower™ NLS-Cas9 Nuclease (MALS verified) （Cat.No: CA9-S5149
）is comparable to that of other competitors
GENPower™ NLS-Cas9 Nuclease is evaluated in an in vitro DNA cleavage assay on a DNA fragment containing the target sequence. The activity of the GENPower™ NLS-Cas9 Nuclease is greater than 90% (QC tested).
The cleavage activity of GENPower™ NLS-Cas9 Nuclease in vitro.
The cleavage activity of GENPower™ NLS-Cas9 Nuclease in cell line.
The cleavage activity of GENPower™ NLS-Cas9 Nuclease in human primary T cells.
High purity verified by SEC-MALS (＞90%)
The purity of NLS-Cas12a Nuclease (Cat. No. CAA-L5149 ) is more than 90% and the molecular weight of this protein is around 135-165 kDa verified by SEC-MALS.
The purity of GENPower™ NLS-Cas9 Nuclease (Cat. No. CA9-S5149) is more than 95% and the molecular weight of this protein is around 145-165 kDa verified by SEC-MALS.
1. Westermann L, Neubauer B, Köttgen M. Nobel Prize 2020 in Chemistry honors CRISPR: a tool for rewriting the code of life. Pflugers Arch. 2021 Jan; 473(1):1-2.
2. Application of CRISPR/Cas9 gene editing in tumor immunotherapy
3. Zaib S, Saleem MA, Khan I. CRISPR-Cas9 Genome Engineering: Trends in Medicine and Health. Mini Rev Med Chem. 2022;22(3):410-421. doi: 10.2174/1389557521666210913112030. PMID: 34517795.