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Genome Editing


Products >  Genome_Editing >  CRISPR_Cas9 >  Resources >  CRISPR_Cas9_Workflow

Design sgRNAs

 

sgRNA design requires the identification of target sites with specific sequence criteria, while also avoiding the potential for off-target effects. Learn about these best practices for designing sgRNAs

   

Transcribe and screen sgRNAs in vitro

 

Quickly transcribe any sgRNA in vitro at high yields, without ligation. Don't waste time delivering ineffective sgRNAs to your cells—test the cleavage efficiencies of individual sgRNAs in vitro before performing gene editing in your target cells

   

Deliver sgRNA and Cas9 to cells

 

There are several options for delivering sgRNAs and Cas9 to your target cells:

For plasmid delivery, our Cas9/sgRNA co-expression vectors allow seamless insertion of your sgRNAs, and express bright fluorescent markers

Use gesicles, which are cell-derived nanovesicles, for efficient delivery of active Cas9 ribonucleoprotein (RNP) complexes to a broad range of cell types with reduced off-target effects and very low cytotoxicity

To prevent genomic integration of Cas9, use this single reagent to transfect your target cells with Cas9 mRNA and sgRNAs without cytotoxic effects


This AAV2-based system for delivery of sgRNAs and Cas9 enables efficient gene editing in difficult-to-transfect cells without genomic integration of Cas9

   

Detect Cas9 protein and confirm gene editing


 

Confirm that Cas9 protein is being expressed in your target cells

Ensure that your cell population contains mutations at your target locus by using a mismatch detection assay that outperforms a CEL-1 based assay

     

Genotype Determination

 

Are there indels on one or both copies of your target gene? Our Cas9/sgRNA-mediated in vitro cleavage reaction can accurately determine your cell's genotype after gene editing

   

Identify
indels

 

Characterize CRISPR/Cas9-induced indels with a simple four-step protocol using the Guide-it Indel Identification Kit

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