Cloning and expression systems for sgRNAs
The Guide-it CRISPR/Cas9 systems are kits for the cloning and expression of single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology. The vectors in these systems simultaneously express Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry (ZsGreen1 and tdTomato versions are available).
The Guide-it CRISPR/Cas9 systems are kits for the cloning and expression of single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology. The vectors in these systems simultaneously express Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry (ZsGreen1 and tdTomato versions are available). Generating a plasmid that expresses a sequence-specific sgRNA with this system is simple: a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex, and the duplexed oligos are inserted into the pre-linearized vector using the included high-efficiency ligation mix. The kit also includes Stellar Competent Cells to ensure high-efficiency transformation.
Overview
- A single vector expresses your target-specific sgRNA from the human U6 promoter; Cas9 nuclease and a bright fluorescent marker (either ZsGreen1 or tdTomato) are expressed from a bidirectional CMV promoter
- tdTomato and ZsGreen1 are exceptionally bright fluorescent proteins, 6x and 2.5x brighter than EGFP, respectively; these markers can be used to monitor transfection efficiency and/or to enrich/isolate transfected cells by flow cytometry
- Simple plasmid construction—design oligos specific to your target sequence and clone into the pre-linearized plasmid using the included ligation reagents and high-efficiency competent cells
- Includes sufficient reagents for construction of 10 different target (sgRNA) expression plasmids
More Information
Applications
- Cloning and expression of a target sgRNA for mammalian genome editing using CRISPR/Cas9 technology
Components
- Guide-it CRISPR/Cas9 System (Green) (Cat. # 632601)
- pGuide-it-ZsGreen1 Vector (Linear)
- Guide-it Ligation Components
- Stellar Competent Cells
- Guide-it CRISPR/Cas9 System (Red) (Cat. # 632602)
- pGuide-it-tdTomato Vector (Linear)
- Guide-it Ligation Components
- Stellar Competent Cells
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
CRISPR/Cas9 information
Choosing sgRNA design tools
Browse a collection of sgRNA design tools for Cas9-based genome editing experiments.
Choosing a target sequence for CRISPR/Cas9 gene editing
Learn how to design sgRNA sequences for successful gene editing.
The CRISPR/Cas9 system for targeted genome editing
Overview of CRISPR/Cas9 system for genome editing.
CRISPR/Cas9 genome editing tools
An overview of tools available for each step in a successful genome editing workflow.
Gene editing technical notes
Delivery of Cas9 and sgRNA to mammalian cells using a variety of innovative tools.
SNP engineering application note
Learn about a simple assay for sensitive detection of single-nucleotide substitutions in bulk-edited or clonal cell populations.
CRISPR/Cas9 gesicles overview
Learn about Guide-it CRISPR/Cas9 Gesicle Production System components and workflow.
CRISPR library screening webinar
Watch this webinar to learn how you can perform genome-wide lentiviral sgRNA screens easily.
Choosing an HDR template format
Watch a webinar on how to choose the right HDR template for knockin experiments.
Guide-it SNP Screening Kit FAQs
Get answers to frequently asked questions and view a video explaining the enzymatic assay.
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