What is Adeno associated virus (AAV)?

AAV is a small non-enveloped single-stranded DNA virus. AAV is dependent on a helper virus for replication. AAV has been engineered as an efficient gene delivery vector (rAAV) for cell and gene therapies. rAAV is not known to cause human disease and has relatively low immunogenicity. AAV genome size is around 4.7 kb, flanked by two 145bp inverted terminal repeats (ITRs).

What are the applications of AAV control viruses?

• AAV eGFP control viruses can be used to monitor the transduction efficiency of your cells 
• AAV eGFP control viruses can be used to select the optimal AAV serotype for your target cell lines
• AAV-eGFP control viruses be used for cell tracking in CNS discovery studies
• AAV empty capsid vectors can be used as negative controls

Tissue Specificity and AAV Serotypes

Variations in AAV capsids lead to different AAV serotypes, which exhibit different transduction tropism for different tissues. You can select the optimal AAV serotype using our AAV serotype testing viral vector panel (SKU AV-SERO-TEST), which contains 7 individual AAV serotypes. The table below summaries the recommended AAV serotypes for different tissues.

Method for Production of AAV Control Viral Vectors

Both the AAV-eGFP and AAV empty capsid viral vectors are prepared using plasmid transient transfection method in HEK293T cells. The AAV-eGFP viral vectors are purified using an iodixanol gradient ultracentrifugation method, while the AAV empty capsid vectors are purified using a chromatographic method.

Custom AAV Packaging Services

If a different AAV serotype or a different amount of viral vector is needed, we can provide a custom AAV packaging service for you. We provide AAV production services covering research grade AAV, AAV for IND enabling preclinical studies, as well as GMP grade AAV for clinical study use. Please contact us at bd@forecytebio.com.

HEK293T cells were transduced with our lentiviral GFP vector at a MOI of 50 (0.5 uL of the premade lentiviral GFP vector, cat# LV-GFP-E9V1). The fluorescence image was taken 72 hrs post transduction.

What Are The applications Using GFP Lentivirus Control Particles?

• The lentiviral GFP vector can be used to monitor the transduction efficiency of your cells 
• The lentiviral GFP vector can be used to optimize the transduction MOI of your specific cell lines
• The lentiviral GFP vector can be used for in vivo cell tracking studies
• The lentiviral GFP vector can be used as a marker control in your gene function studies

Preparation Of our Lentiviral GFP Vectors

The lentiviral GFP vectors are produced using a plasmid transient transfection method by co-transfecting pGag-pol, pRev, pVSV-G and the lentiviral transfer plasmid, pEF1a-GFP-Puro, into the HEK293T cells. The harvested viral vector supernatant is purified and concentrated using a ultracentrifugation method. The final lentiviral vectors will be resuspended in phosphate buffered saline (PBS).

What Is MOI and What Is The Optimal MOI For My Cells?

MOI stands for multiplicity of infection, the number of transducing lentiviral vector particles to each cell. The MOI used for lentivirus transduction of different types of cells varies, ranging from 3-100. For efficient gene delivery into different cell types, the optimal MOI can be tested using our lentiviral GFP vectors.

Need Custom Lentiviral Vector Packaging?

We can provide a custom lentiviral vector production for you if a different scale or different reporter control /other GOIs is needed. We provide lentiviral vector production services covering research grade lentiviral vectors, lentiviral vectors for IND enabling preclinical studies, as well as GMP grade lentiviral vectors for clinical study use. Please contact us at bd@forecytebio.com.

Lentivirus Resources

GFP Lentivirus Control Data Sheet

Lentivirus Transduction Protocol