AVV Transduction

TaKaRa Bio

1) AAV Packaging

293T Cell Line

• Suitable for adeno-associated virus (AAV) packaging
• Supports the high level expression of viral proteins in high titers

AAV Helper-Free Packaging Plasmid Set

• et of two plasmids (pRC-mi342 and pHelper) designed to get a high titer AAV2
• AAV2 particles are generated without the use of a helper virus and are widely used in in vivo studies

 

2) AAV Transduction Tools

AAVpro Extraction Solution

• Is an extraction buffer that increase AAV yield at least 3-fold
• It provides a simple method for AAV particle isolation only by centrifugation
• Can be used for any AAV serotype

 

AAVpro Titration Kit

• Is a qPCR titration kit
• Contains all the components necessary for extraction of AAV particles from AAV producing cells and real-time PCR analysis of a viral genomic sequence
• The workflow lasts approximately 2,5 hours and the titration is with high sensitivity

 

AAVpro Purification Kit

• Uses a column based purification system to quickly and easily purify AAV serotype 2 viral particles without need for ultracentrifugation.
• The faster way unlike methods such as cesium chloride concentration gradient ultracentrifugation or iodixanol ultracentrifugation
• This methods are commonly used to purify AAV particles, but such methods are time consuming and require careful technique to obtain high yields

 

3) AAV Vector Systems

AAVpro Helper Free System (AAV2)

• Kit for the preparation of serotype 2 AAV particles without the use of the helper virus
• Includes three plasmids encoding the factors necessary to prepare recombinant AAV particles and reagents for extracting AAV particles from producer cells

 

pAAV ZsGreen1 Vector

• Can be used as a positive control to monitor AAV particle production using the AAVpro Helper Free System (AAV2)
• The expression of the green fluorescent protein ZsGreen can be used to monitor the efficiency of the transfection and transduction

 

AAV Tet-Inducible Expression System

• Tet-One Systems have all the components for tetracycline inducible expression on a single vector
• Tet-On 3G transactivator is expressed in from the human phosphoglycerate kinase 1 (PGK) promoter
• Cloned gene of interest is expressed from the P_TRE3GS promoter
• This all in one design has been shown to result in up to 25 000 fold induction

 

AAV Cre Recombinase System

• Kit for preparation of AAV2 particles to deliver a Cre recombinase gene under the control of the CMV promoter
• It can be used in transduction of target cells containing a floxed DNA sequence with the AAV2-CRE Recombinase particles
• Results in recombination between the loxP sites or it can be used for generating the transgenic mice

 

AAV2-CRISPR/Cas9 System

• Viral delivery of the Cas9/sgRNA complex allows to edit hard to transfect cells
• The Cas9 gene is splitted between two plasmids with 1,6 kb region of homology which produce the full length Cas9 gene
• Expression of a human microRNA miR-342 from the packaging plasmid significantly increases AAV titer

 

AAV-LacZ Particles

• Kit to prepare AAV2 particles for delivery of the lacZ reporter gene to mammalian cells under the control of the CMV promoter
• Can be prepared without a helper virus
• The lacZ gene encodes the commonly used reporter, enzyme beta-galactosidase

 

Documents and resources

For more information visit producer's website.