Our research-grade retrovirus production service lets you choose from multiple expression plasmid vectors depending on the expression type you desire. Stable-cell-line and iPS-cell generation are also available.

Retroviral vector production

1. Plasmid construction and preparation

• Cloning of your gene of interest (GOI) into the expression plasmid vector of your choice
• Large-scale preparation of plasmid DNA for retroviral vector production

2. Producer cell production (optional)

• Recommended when performing retroviral vector transduction to human PBMCs.

3. Retroviral vector production

• Transfection of 293T cells with plasmid DNA (e.g., pDON-5-GOI and packaging plasmid DNA)
• Transient expression of retroviral vector and virus collection
• Viral titration by real-time PCR

4. Large-scale retroviral vector production

• Production of retroviral vector at the requested scale

5. Stable-cell-line generation (optional)

• Transfection of target cell line using the retroviral vector
• Antibiotic selection of cells expressing GOI
• Confirmation of GOI expression by real-time PCR

Our research-grade lentivirus production service uses Takara Bio’s proprietary transient lentiviral vector production system. Large-scale preparation and purification by ultracentrifugation are also available.

Lentiviral vector production

1. Plasmid construction and preparation

• Cloning of your gene of interest (GOI) into expression plasmid DNA (pLVSIN-GOI)
• Large-scale preparation of plasmid DNA for lentiviral vector production

2. Lentiviral vector production

• Transfection of 293T cells with plasmid DNA (pLVSIN-GOI, packaging plasmid DNA)
• Transient expression of lentiviral vector and virus collection
• Viral titration by real-time PCR

3. Large-scale lentiviral vector production and purification

• Production of lentiviral vector at the requested scale
• Purification by ultracentrifugation, etc.

4. Stable-cell-line generation (optional)

• Transfection of target cell line using the lentiviral vector
• Antibiotic selection of cells expressing GOI
• Confirmation of GOI expression by real-time PCR

Features of our research-grade AAV production service include:

• Safer, helper-independent AAV vector with AAVpro Helper Free System
• Highly purified AAV vector with Takara Bio’s proprietary extraction method
• Multiple choices of purification method to suit your application
• Scale-up development for future commercial manufacturing is also available

AAV vector production

1. Plasmid construction and preparation

• Cloning of your gene of interest (GOI) into expression plasmid DNA (pAAV-GOI)
• Large-scale preparation of plasmid DNA for AAV vector production

2. AAV vector production (crude)

• Transfection of three plasmids (pAAV-GOI, pRC, and pHelper) into 293T cells
• Transient expression of AAV vector

3a. AAV vector purification from cell pellet

• Collection of AAV vector from cell pellet
• Purification with AAVpro Purification Kit (All Serotypes)

or 

3b. AAV vector purification from cell pellet and supernatant

• Collection of AAV vector from cell pellet and supernatant
• Purification by affinity chromatography

4. AAV vector purification with ultracentrifugation (optional)

• Additional purification with cesium chloride gradient centrifugation

Our research-grade adenovirus production service produces high titer, high purity virus for in vivo administration. The adenoviral vector is made from producer cells derived from a single clone.

Adenoviral vector production

1. Cosmid vector construction

• Cloning of your gene of interest (GOI) into cosmid vector
• Large-scale preparation of cosmid vector for adenoviral vector production

2. Clonalized adenoviral vector production

• Transfection of 293 cells with the adenovirus DNA containing GOI digested from cosmid vector
• Collection of adenoviral vector from cell supernatant (“full-length transfer method”)
• Viral titration by hexon detection and RCA contamination testing
• Viral titration by TCID50 is also available

3. Adenoviral vector purification (for large-scale [>1 L] only)

• Purification by cesium chloride gradient centrifugation
• Viral titration by hexon detection and RCA contamination testing
• Viral titration by TCID50 is also available

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