...Unlike lentivirus, AAV does not possess the ability to replicate itself and depends on helper viruses like...like adenovirus (Ad) or herpes simplex virus to facilitate efficient replication. The most common production...transfection reagents and keep the cells happy. Harvesting virus Once you have transfected your cells, wait 2–5 days...to an overestimate of transducible units in the virus preparation.  rAAVs  Now your AAV virions are ready...detection and quantification of adeno-associated virus serotype 2-derived inverted terminal repeat sequences...Characterization of a recombinant adeno-associated virus type 2 Reference Standard Material. Human Gene Therapy...into the culture medium during adeno-associated virus manufacturing. Human Gene Therapy, 21(10), Article...

... a contaminant of adenovirus preparations. It is an icosahedral, non-enveloped virus that carries a small...Parvoviridae and requires co-infection of helper viruses like adenovirus for productive infection. AAV vectors are...directly to the cells that need them. Adeno-associated virus (AAV) – based delivery techniques may hold the key...delivery tools: Unlike most naturally occurring viruses used in clinical trials, wild type AAV is not associated...These pre-existing antibodies can interfere with virus-cell interactions and seriously hinder gene therapy...., et al. "Immune responses to adeno-associated virus and its recombinant vectors." Gene therapy 10.11... adeno associated virus...

...interact with the virus. As a result, we identified 332 human proteins that the virus interacts with in...on BioRxiv describing a comprehensive SARS-CoV-2 virus-human interaction network in which we expressed ...in cells. Because viruses rely on the help of human proteins in their lifecycle, identifying these potential...strategy to throw a therapeutic monkeywrench into the virus’ hijacking of human cells. Even better, more than...interactors we found in our map have an impact on virus replication. While we and our collaborators are ...Manon Eckhardt and Melanie Brewer from the QBI Coronavirus Research Group at UCSF. It’s been only a few ...scientists from more than 20 labs to form the QBI Coronavirus Research Group (QCRG) to do just this.  In under...

...University, contributed to this post. Adeno-associated viruses (AAVs) make fantastic gene delivery vehicles for...gene knockouts using recombinant adeno‐associated viruses." Nucleic acids research 32.1 (2004): e3-e3. PubMed... et al. "Directed evolution of adeno-associated virus for enhanced gene delivery and gene targeting in...Schofield. "Genome engineering using adeno-associated virus (AAV)." Chromosomal Mutagenesis. Humana Press, New... adeno associated virus...

...After months of input and polling, Blugene's new virus friend finally has a name! We thank you for your...In December, we introduced you to Blugene’s new virus friend (they/them/theirs). But, they didn’t have...

...housekeeping activities.       Viruses and biosensors So how do viruses factor into all of these diverse...review the basics of biosensors and how to use viruses to set up these systems. Components of a biosensor...This specificity is feasible partially because of viruses—keep reading to find out how!   Neurotransmitters...and downstream monitoring. Nervous about making a virus for your biosensor? Don’t be! We have you covered...resources and safety tips. Still worried? We make virus with some of our most popular biosensors already...deliver your sensor, whether you choose lentivirus, gamma retrovirus, or AAV as your vehicle, we’ve got you...For one, many viral platforms, such as AAV or retrovirus, can either permanently integrate and/or express...

...generating your lentivirus, you’ll want to infect your cells with the virus–the right amount of virus, that is...to make your virus. If the library happens to have a specific protocol for producing virus, I recommend...received your pre-made library/produced your lentivirus. Since viruses are sensitive to freeze/thaw cycles, we...production steps before you can start transducing your virus. Let’s start there!     Figure 1: Flowchart...number of freeze-thaw cycles needed when using the virus. A small aliquot is used to determine functional...HEK cells allows production of lentivirus. Purification of this lentivirus results in the pooled library...our protocol on Lentivirus Production provides a general approach for producing lentivirus in Lenti-X 293T...

...pseudorabies virus type 1, vesicular stomatitis virus (VSV; Beier et al., 2011), adenovirus, and adeno-...origins of these methods are naturally occurring viruses that infect, persist, and migrate within neurons...synaptic connections. Most famous is the Rabies Virus (RABV) which moves retrogradely from a peripheral...review see Dietzschold et al., 2008). Herpes Simplex Virus (HSV) also migrates through neurons where it can...connections (Ugolini et al., 1989). When using these viruses to mark neuronal connectivity a large majority ...overtime due to the continued replication of the virus. While this can be useful, if too many neurons are...Anderson, 2011). In the past two decades, several virus species have been modified and used as tools for...

...typically DNA viruses, such as adeno-associated virus, herpesviruses, hepatitis B virus, adenoviruses, and human...human papillomavirus (HPV), to name a few (Desfarges & Ciuffi, 2012). One family of viruses, retroviridae...genome isn’t uncommon. Many naturally occurring viruses occasionally integrate their genomes into those...plasmid typically encodes the vesicular stomatitis virus G protein (VSV-G), which facilitates viral entry...Integration and Consequences on Host Gene Expression. Viruses: Essential Agents of Life, 147–175. https://doi.org... & Varmus, H. E. (1996). Human immunodeficiency virus type 1 integrase mutants retain in vitro integrase...integrase-deficient lentiviruses (IDLVs). As the name suggests, these lentiviruses lack a functioning ...

...of Gene Delivery to the Kidney by Adenovirus, Adeno-Associated Virus, and Lentiviral Vectors After Intravenous.... M., & Sweeney, H. L. (2008). Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer...vivo-directed evolution of a new adeno-associated virus for therapeutic outer retinal gene delivery from... J., & Chiorini, J. A. (2001). Adeno-Associated Virus Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid...2016). An essential receptor for adeno-associated virus infection. Nature, 530(7588), 108–112. https://doi.org... Zabner, J. (2001). Binding of adeno-associated virus type 5 to 2,3-linked sialic acid is required for...Stable Pancreatic Gene Transfer by Adeno-Associated Virus Vectors via Different Routes. Diabetes, 55(4), 875...

... endo-lysosomal trafficking of adeno-associated virus/phage." Journal of Biological Chemistry (2012): ...into the culture medium during adeno-associated virus manufacturing." Human gene therapy21.10 (2010): ...phase, but not the production phase, improves adenovirus production." Journal of biotechnology 157.1 (...

...production is more complex and the presence of helper virus contaminants have slowed the use of this system....which contains the adenoviral genes necessary for virus production. For detailed instructions on how to ... together ~3 days post infection. The amplified virus can be harvested once roughly 50% of the infected...amplification should result in a 10-100-fold increase in virus. Purification (2 days) - Purification is required... (280nm) absorbance. The ratio for the purified virus should be ~1.3 Quantitative PCR: the number of...Additional Resources on the Addgene Blog Using virus in your research - a primer for beginners Adenoviral... are based on Adenovirus type 5. Three generations of adenoviral vectors The adenovirus virion is a non-enveloped...

...indicators work, their general applications, and why viruses are essential to their use as biological tools....transiently and for genomic integration. Lab-engineered viruses will package your genetic cargo in lieu of their...deposit this cargo upon host infection. Packaging viruses also have various serotypes – the surface proteins...tool that swaps capsid proteins from different viruses to optimize them for targeting your cell type of...essential. If you want to learn more about how viruses can be used for chemigenetics, biosensors, and ...Addgene.org Addgene’s Viral Services Addgene’s Virus Protocols Addgene’s Chemogenetics Guide Resources...

...Please note that the generation used to package the virus does not change the generation of the transfer vector...What envelope glycoprotein will be expressed on virus particles? A5: This depends on the envelope plasmid... choice of envelope dictates the tropism of the virus.  VSV-G is very common due to its broad host range...replication-deficient meaning they can be used to create virus that is capable of infecting target cells, but cannot...contain the minimal number of HIV genes required for virus production (3 or 4). Third-generation vectors contain...the potential generation of replication competent virus [usually HIV-1]; and 2) the potential for oncogenesis... often: Q1: Are retroviruses and lentiviruses the same thing? Can I package lentivirus with retroviral...

...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging ...the transgene. When a cell is transduced by both viruses, if the AAV genomes encoding part A and part B ...Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27:1–12 . https...Fragmented Genome Reassembly of Adeno-associated Virus. Molecular Therapy 18:6–8 . https://doi.org/10.1038...

...plasmid which contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains...that a gene of interest can be packaged into the virus instead.  Next let’s take a closer look at the rep...Perry WL III, Strohl WR (2017) Adeno-Associated Virus (AAV) as a Vector for Gene Therapy. BioDrugs 31:...  Maurer AC, Weitzman MD (2020) Adeno-Associated Virus Genome Interactions Important for Vector Production...

... surface. But detection of the virus costs time and money, and viruses at low concentrations often escape...tailored for the detection of other norovirus strains and even other viruses by modifying the sequence of the...switches to detect Zika virus (available at Addgene). The plasmids for detecting norovirus will be coming soon...antibodies--attached to magnetic beads, allowing the virus to be easily concentrated. Viral RNA is then amplified...presence of the norovirus. While this assay is specific to a particular strain of norovirus, it can be tailored...both the stomach flu and food-borne illnesses, noroviruses can infect millions annually. Because it is highly...analyst is necessary. How does this assay work? The norovirus is enriched with synbodies--or synthetic antibodies...

...pseudotype the spike protein onto another virus. With the pseudovirus, scientists can study the mechanisms ...protein, variants, and pseudotyping The SARS-CoV-2 virus depends on its spike protein to bind ACE2, the host...vaccines and diagnostics (Hsieh et al., 2020). As the virus spreads around the world, the spike protein has ... pseudotyping, visit the SARS-CoV-2 Pseudotyped Virus collection page. SARS-CoV-2 detection tools There...for SARS-CoV-2 researchers to investigate how the virus interacts with human cells. Addgene’s mammalian ...SARS-CoV-2 Spike Protein for Neutralization Assays. Viruses 12:513. https://doi.org/10.3390/v12050513 Hsieh...that can be incorporated into pseudoviruses. The lab used these pseudoviruses to determine how effective ...

...also stops viral vectors, such as adeno-associated virus (AAV), from reaching their targets. The AAV9 serotype...labs determined LY6A was a receptor used by PHP.B viruses. Creating PHP.B, a neurotropic AAV9 variant PHP.B...injected with PHP.eB, an enhanced version of PHP.B, virus still reached the brain endothelial cells of these...contributed to the unintentionally selection of a virus with mouse-strain dependent performance. This surreptitious...Intravenous administration of the adeno-associated virus-PHP. B capsid fails to upregulate transduction efficiency...Addgene or check out protocols for packaging your own virus ...members are linked to viral susceptibility (mouse adenovirus in mouse; HIV1, West Nile, and Influenza in humans...

...= Copies/uL V = Volume of virus in reaction mix D = Dilution factor of virus Most droplet readers have...quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used to confirm that ...to titrate AAV by ddPCR begins with diluting the virus. It is important to note that the dynamic range ...1012 to 1013 GC/mL, you must serially dilute your virus before adding your sample to a mastermix. At Addgene...should avoid pipetting too quickly when making your virus dilutions. This will reduce aerosols that could ...