CRISPR/Cas9 Do-It-Yourself Tools
Cas9 (CRISPR associated protein 9) is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) adaptive immunity system in Streptococcus pyogenes. S. Pyogenes utilizes Cas9 to remember and later probe and cleave foreign DNA, such as invading bacteriophage DNA or plasmid DNA. When the DNA substrate is complementary to the guide RNA (gRNA), Cas9 cleaves the invading DNA.
This ability to make site-directed double strand breaks in DNA has led to the use of Cas9 protein as a genome engineering tool. As an RNA-guided DNA endonuclease, Cas9 can be directed to specific target cleavage sites by altering the guide RNA sequence. The nuclease-deactivated form of dCas9 further provides a versatile tool for biomedical research and therapeutics.
Stratech Scientific offers a comprehensive range of CRISPR/Cas9 related enzymes, proteins, antibodies and kits for genome engineering from expert suppliers NovateinBio and System Bioscience (SBI).These products include Cas9 and gRNA delivery systems in a flexible range of formats, homologous recombination donor vectors, and AAVS1 Safe Harbor targeting, as well as Cas9 transfection ready proteins and screening tools—we can help bring your genome engineering projects to life.
Cas9 Transfection Ready Protein
Compared to transfection with Cas9-expressing plasmid, delivery of purified Cas9 protein directly into cells by electroporation is relatively straightforward and may reduce the risk of issues associated with plasmid-based transfection, such as random integration of plasmid into the host genome.
The presence of a nuclear localization signal (NLS) localizes the Cas9 RNP complex to the nucleus immediately upon entering the cell with the need for transcription or translation compared with mRNA or plasmid systems.
Additionally, the Cas9 RNP complex is rapidly cleared from the cells minimizing the chance of off-target cleavage when compared to other systems. There is no need for transcription or translation processes.
Stratech offers a range of transfection ready recombinant and engineered Cas9 nucleases and nickases.
NLS-Cas9-NLS, NLS-dCas9-NLS, NLS-Cas9-EGFP, NLS-Cas9(D10A)-2NLS nickase
CRISPR/Cas9 plasmids & guide RNA Delivery tools
With a wealth of options for delivering Cas9 and gRNA, Stratech supports a full range of genome engineering applications. Deliver Cas9 and gRNA via mRNA, lentivector, or AAV, or use purified Cas9 protein, and then choose from our all-in-one systems that deliver Cas9 and gRNA on a single vector or two-component systems for separate introduction of Cas9 and gRNA.
Multiplex Gene Cloning
Take your CRISPR/Cas9-based genome engineering experiments to the next level with Multiplex gRNA Cloning Products. Easy-to-use and efficient, they streamline the workflow for making multiple changes to a genome by delivering multiple gRNAs on a single vector.
PrecisionX Multiplex gRNA Cloning Kit
Multiplex gRNA Kit + Cas9 Nickase: EF1-T7
Multiplex gRNA Kit + Cas9 Nickase: CAG-T7
Screening Tools for CRISPR/Cas9
Easily verify Cas9 mRNA and protein expression with Cas9 antibody and ready-to-go Cas9 RT-PCR Primers.
Designed to detect wild-type Cas9, Nickase (D10A, H840A), and catalytically dead Cas9 (dCas9), our Cas9 Antibody (Clone 7A9) is an IgG1 mouse monoclonal antibody that is Protein G affinity-purified and works well in range of Cas9 detection assays, including Western blot analysis, immunofluorescence, and immunoprecipitation.
Confirm Cas9 mRNA expression with ready-to-go Cas9 RT-PCR Primer Set.
The amplicon for Primer Set 1 is 219bp and amplicon for Primer Set 2 is 122bp. These primer sets allow for detection of Cas9 nuclease, Cas9 Nickase and Cas9 double mutant at messenger level. The primer sets are compatible with Cas9 mRNA expression from any SBI Cas9 SmartNuclease construct and vectors.
The sgRNA Screening Kit is designed to quickly assess in vitro the potential efficiency of a sgRNA sequence in vivo
- sgRNA Screening Kit
Cas9 detection antibodies
- Anti-CRISPR/CAS9 Monoclonal Antibody
- Anti-CRISPR/CAS9 Polyclonal Antibody
The CRISPR/Cas9 Protein ELISA Kit (Colorimetric) is designed for measuring Cas9 level in whole cell extracts or purified Cas9 proteins from various cells and tissues. CRISPR/Cas9 Protein ELISA Kit (Colorimetric)
Homologous Recombination Targeting vectors
Taking advantage of the cell’s double-strand break (DSB) repair system, homologous recombination (HR) targeting vectors (also known as HR donor vectors) can increase the efficiency of gene knock-outs, and are essential for gene knock-ins, edits, and fusions. HR Targeting Vectors are all that and more, with features that can make it easier to find successful clones, minimize the amount of vector DNA left in the genome, minimize off-target integration, and more.
- Basic HR Targeting Vector
- Gene Knock-Out HR GFP Targeting Vector
- Gene Knock-Out HR RFP Targeting Vector
AAVS1 Safe Harbor Targeting
With HR donor vectors that already contain AAVS1 homology arms, these products streamline genome engineering at the AAVS1 Safe Harbor Site. Create isogenic cell lines, reporter cell lines, Cas9-expressing cell lines, and more, and take advantage of the consistent, robust transgene expression possible at the AAVS1 site.
AAVS1 Safe Harbor Targeting Vector 2.0 – GOI Knock-in Donor
AAVS1 Safe Harbor Targeting Vector 2.0 – All-Purpose Donor Complete Kit with Cas9 -gRNA Targeting Vector and Junction PCR Primer Mix to confirm AAVS1 integration site
AAVS1 Safe Harbor Targeting Vector 2.0 – GOI Knock-in Donor, Complete Kit with Cas9 gRNA Targeting Vector and Junction PCR Primer Mix
CRISPR/Cas9 Services
Through our partner, SBI, we also offer the below services
CRISPR/Cas9 gRNA and HR Donor Cloning
With this solution, you will be able to simplify both the design and cloning of the gRNA or the homologous recombination donor constructions. Using the latest technology it is possible to simplify the process down to key elements and ensures successful execution on any project. You will be able to save yourself both time and effort as you allow experts to complete intricate processes.
You can choose either gRNA and HR donor design or select the Cell Line Engineering Service.
CRISPR/Cas9 Cell Line Engineering
Using this service it is possible to ensure that CRISPR/Cas9 cell line engineering is reliable and easy under the guidance of an experienced team. Deliveries are rapid and full confidentiality is guaranteed.
Using the latest technology it is possible to simplify the process of genome engineering and ensure that time can be saved throughout the process. It is also possible to avoid some of the common pitfalls and mistakes while guaranteeing that project requirement are met.
