Takara Cloning and expression systems for sgRNAs

Takara Guide-it CRISPR/Cas9 Systems for sgRNA Cloning and Expression

The Guide-it CRISPR/Cas9 systems are kits for cloning and expression of single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology.
These vectors simultaneously express Cas9 nuclease, a target-specific sgRNA, and a bright fluorescent protein (ZsGreen1 or tdTomato) for monitoring transfection efficiency or isolating transfected cells by flow cytometry.

Generating a plasmid that expresses a sequence-specific sgRNA is simple:
a pair of user-provided oligos corresponding to the target genomic sequence are annealed to form a duplex, then inserted into the pre-linearized vector using the included high-efficiency ligation mix.
The kit also includes Stellar Competent Cells for high-efficiency transformation.

Overview

• Single vector expresses target-specific sgRNA from the human U6 promoter
• Cas9 nuclease and a fluorescent marker (ZsGreen1 or tdTomato) expressed from a bidirectional CMV promoter
• tdTomato and ZsGreen1 are 6× and 2.5× brighter than EGFP, respectively
• Bright markers enable monitoring of transfection efficiency and/or enrichment of transfected cells
• Simple plasmid construction using included ligation reagents and competent cells
• Sufficient reagents for constructing up to 10 sgRNA expression plasmids

Applications

• Cloning and expression of target sgRNAs for mammalian genome editing using CRISPR/Cas9

Components

Additional Product Information

Please refer to the product’s Certificate of Analysis for details on storage conditions, components, and technical specifications.
Kit Components Lists and Certificates of Analysis are available under the Documents tab.