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Products >  Viral_Transduction >  AAV_Vector_Systems >  CRISPR_Cas9

Helper Virus-Free Preparation of AAV particles to deliver Cas9 and sgRNA

The AAVpro CRISPR/Cas9 Helper Free Systems are used for helper virus-free preparation of AAV2 particles to deliver expression cassettes for Cas9 and a user-defined single guide RNA (sgRNA) to mammalian cells. This two-vector system overcomes the size restrictions of the AAV genome to deliver the large Cas9 gene to target cells, allowing for efficient genome modification in a wide variety of mammalian cells in vitro. High-titer AAV2 particles can be prepared without a helper virus by simply transfecting HEK 293T cells with the included plasmids that encode all of the factors necessary to prepare recombinant AAV particles. The AAVpro CRISPR/Cas9 Helper Free System is a complete system, containing all the reagents necessary to clone the sgRNA into the expression plasmid and prepare AAV particles. AAV Extraction Solution is also included for efficient AAV2 viral particle isolation.

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Viral delivery of the Cas9 gene and a sgRNA allows for genome editing in hard-to-transfect mammalian cells, including proliferating and non-proliferating cells
  • AAV delivery of Cas9 precludes genomic integration and persistent expression of Cas9, reducing off-target effects 
  • The Cas9 gene is split between pAAV-Guide-it-Up and pAAV-Guide-it-Down plasmids with a 1.6-kb region of homology; this homologous region results in recombination in target cells, producing a full-length Cas9 gene 
  • pGuide-it-Down plasmid is provided prelinearized, facilitating single-step cloning of sgRNA with reduced background due to re-ligation of plasmid  
  • Expression of human microRNA miR-342 from the packaging plasmid significantly increases AAV titer 

Applications

  • AAV-based delivery of a user-defined sgRNA and Cas9 for mammalian genome editing using CRISPR/Cas9 technology

Components

AAVpro CRISPR/Cas9 Helper Free System (AAV2) (Cat. No. 632608)

  • AAVpro CRISPR/Cas9 Vector Set
  • Guide-it Ligation Components
  • Stellar Competent Cells
  • pRC2-mi342 Vector
  • pHelper Vector
  • AAVpro Extraction Solution

AAVpro CRISPR/Cas9 Vector System (Cat. No. 632609)

  • AAVpro CRISPR/Cas9 Vector Set
  • Guide-it Ligation Components
  • Stellar Competent Cells

Additional 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.

 
 
Products
Cat. # Product Contents Size Price License Units Select
632608 AAVpro® CRISPR/Cas9 Helper Free System (AAV2) 1 System $894.00 License Statements
632609 AAVpro® CRISPR/Cas9 Vector System 1 System $502.00 License Statements
 

 

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631448 Guide-it™ Mutation Detection Kit 25 Rxns $188.00
 
 
 

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