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  1. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    ...pathogenic Ebola virus belongs to the group of nonsegmented negative sense RNA viruses, along with other...other viruses that cause disease in humans such as measles, mumps, and rabies. Research on Ebola virus has... sense RNA viruses (reviewed in 2, 3). Other important components of the Zaire ebola virus minigenome ...respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue...transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems....replication competent virus-like particles and beyond: reverse genetics systems for filoviruses and other negative...part, due to the necessity for working with this virus under the highest biosafety level conditions, BSL...

  2. Viral Vectors 101: Viruses as Biological Tools

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    ...types of viruses used as biological tools.   Viruses as biological tools In nature, viruses infect cells...exactly which type of virus to use. Types of viruses There are four main types of viruses used in mammalian...research: 𝛄-retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV). While each virus allows ...experiment. Retro and lenti viruses Retroviruses are a family of viruses that stably integrate their ...features that make these familiar viruses such a pain is exactly why viruses are such wonderful scientific... means viruses can be engineered to be cell type specific. Once inside the host cell, the virus unleashes...isn’t enough to ensure the virus can replicate and spread. Instead, viruses replicate by hijacking the...

  3. 5 Tips for Troubleshooting Viral Transductions

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    ...which the virus is released, a more concentrated virus solution can be obtained. 4. Enhancing virus-cell contact... Addgene Blog Learn All about Adenovirus Associated Virus New to Virus? Check out Our Primer for Beginners...transfected and produced infectious virus, they may get infected with this virus, and therefore, will be selectable...necessarily indicate the production of virus. Alternatively, if the virus encodes a fluorescent protein, the...Regardless of the type of virus you’re working with, there’s a lot that goes into virus design, production, ... freeze-thaw cycles on hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with...Viral Vector Guide Pages Find Virus Protocols Order Virus ...

  4. Viral Vectors 101: What is a Virus (and a Viral Vector)?

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    ...talking about viruses, here’s a quick tip on terminology. “Virus” refers to a single type of virus, not the... as viruses) are named after the viruses they’re derived from: AAVs, lentivirus, gamma retrovirus, and...biology tools, a very useful trait. Virus structure While all viruses have a genome inside a protein shell...envelope. Viruses without an envelope are referred to as naked or non-enveloped, while viruses with an ... called enveloped viruses (Figure 1). Like bacteria, only a tiny fraction of viruses are pathogenic (Balloux...2017). Figure 1: A non-enveloped virus (left) and an enveloped virus (right). Created using biorender....Taylor, M. W. (2014). What Is a Virus? In M. W. Taylor (Ed.), Viruses and Man: A History of Interactions...

  5. Viral Vectors 101: Virus Safety

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    ... derived from viruses that can infect people, cells, and animals. Some of these viruses are completely...While the virus can still enter (and thus infect) cells, this safety feature prevents the virus from replicating... concern with virus safety is the disease-risk of what is being integrated by the virus. If an oncogene...infected by a lab-grown virus, it's important to understand the risks of the specific virus you’re working with...treatment and/or monitoring. Virus-specific concerns   In nature, some viruses are more dangerous than others...true for types of viruses generated in the lab. The four most common types of viruses grown in the lab ...integrating viruses – they physically deposit their cargo into the host’s DNA. These viruses also have ...

  6. Viral Vectors 101: Types of viruses

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    ...commonly used lab viruses: gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus. Gamma-retrovirus...used for either type of retrovirus. Adeno-associated virus Adeno-associated virus (AAV) is a small, single-stranded... Choosing a virus Now that we’ve reviewed the common viruses used in biological research, you might ...An Overview of the Application of Viruses to Biotechnology. Viruses. 2021 Oct 14;13(10):2073. DOI: 10.3390...about a specific virus? We have you covered with guides on lentivirus, retrovirus, adenovirus, and AAV. If...But these viruses are roomy: some adenoviruses pack up to ~35 kb of space for cargo! Adenovirus is an effective...Gamma-retrovirus Retroviruses are single-stranded RNA viruses that use reverse transcriptase to synthesize complementary...

  7. Production of Virus in Insect Versus Mammalian Cells

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    ...to grow. The virus Insect cell systems are used to produce baculovirus, a family of viruses that specifically...custom viruses are too high for your budget. In those cases, you’ll need to produce your viruses in-house...type of cells that will be used to make the virus. Viruses can be produced in many different cell types...these viruses. The cells are also easy to cultivate, fast-growing, and widely available. The viruses Lentiviruses...producing virus within 24 to 48 hours. Harvesting timepoints will depend on your specific virus, so this...24 hours. Processing the virus-containing supernatant will depend on the virus type. AAV is typically purified... of virus, called P1 (or sometimes P0), infecting new Sf9 cells, and getting an amplified P2 virus. This...

  8. Viral Vectors 101: Transductions

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    ... the production cell line (depending on the virus), virus can be collected for the transduction step. ...swap out your virus type to achieve different biological outcomes. For example, some viruses integrate into...to be injected with a virus, but enables tissue-specific targeting of that virus, which can’t be easily...specific viruses available to choose from. And remember, we have a selection of ready-made viruses as well...Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene Viral Services...the mechanisms vary. Why transduce? So why use a virus to deliver nucleic acids? Sounds sort of dangerous...dangerous at first suggestion! The good news is that viruses are very valuable biological tools, and with the...

  9. Viral Vectors 101: Pseudotyping

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    ...only done on enveloped viruses such as lentivirus, retrovirus, and rabies virus. How to pseudotype To ... allows these viruses to infect neurons. Besides altering the host tropism of your virus, there are many... the virus and is less affected.  Altering sensitivity to serum. Different pseudotyped viruses are differentially...production of vesicular stomatitis virus, HIV-2, and human foamy virus in cell lines expressing galactosyl...SARS-CoV-2 virus. Scientists have been using similar pseudotyping methods with other viruses in a BSL-2... because of rabies virus’s toxicity, it made more sense to pseudotype another virus with rabies glycoprotein...the cell body. Rabies virus and the EnvA-TVA system Another way to use rabies virus is to delete the glycoprotein...

  10. Viral Vectors 101: Viral Vector Elements

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    ...the viruses discussed above (gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV...here! Need Virus? Check out Addgene's Viral Service! Types of viral vectors Of the many viruses out there... by virus and then by function. Lentivirus/gamma-retrovirus Lentiviruses are a class of retrovirus (so...will discuss the gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV). The components...there are many shared components across viruses (AAV, lentivirus, etc.) and their functions are more intuitive... 1 - Viral vector components of retrovirus  Adeno-associated virus AAV’s elements are quite different... viral elements. Gene expression Adenovirus is a linear, dsDNA virus naturally. It has ITRs, just like...

  11. Using Addgene's New Viral Service for Your Research

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    ...producing virus from select vectors in the repository yourself, you can now order ready-to-use virus directly...starting work with virus. You can find all of our viral vector blog posts here. If you have virus expertise you...as well as expanded virus protocols, information on how we produce and test virus, and quality control...genome-wide screen, we’ve got viruses to help you out. Of course, lentiviruses are used in experiments other...that are currently available) and adeno associated virus (AAV, primarily chemogenetics tools for now but ... to accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the ... help them share and distribute other reagents. Virus distribution came up time and time again with many...

  12. Viral Vectors 101: The Retroviral Lifecycle

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    ...essentially the same for viruses and viral vectors. Key to the wild-type virus’ reproductive cycle is the...of the Bryan strain of Rous sarcoma virus: purification of the virus and isolation of the nucleic acid....(1974). Infectious rous sarcoma virus and reticuloendotheliosis virus DNAs. Journal of Virology, 14(5)...lab because it is, after all, among the more virulent viruses (it attacks the immune system directly). It...cells (i.e. transferring DNA), yet retroviruses are RNA-containing viruses  (Robinson et al., 1965)! You may...odder when you consider that out of the 6 x 106 virus-like particles that we come into contact with every... the fact that the virus must have its structural proteins expressed from a reverse-transcribed piece ...

  13. Adapting Toehold Switches to Detect Zika Virus

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    ...the Zika virus genome with Addgene to facilitate rapid sharing of these tools with the Zika virus research...American-lineage of the virus. For the first time, we have also demonstrated our technology on active virus, as well ...detect the virus in a clinically relevant setting. Molecular diagnostics for the Zika virus predominantly...background First identified in 1947 in Uganda, the Zika virus had received little attention and, for the most ...in January with the report of an outbreak of the virus in Brazil that was correlated with greater rates...albopictus) and, aided by international travel, the Zika virus is expected to expand into heavily populated regions...resistance and strain-specific diagnostics for the Ebola virus. Our original proof-of-concept work was exciting...

  14. Viral Vectors 101: Viral Applications

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    ... recommendations for putting viruses to work. Viral applications Viruses work by infecting a host cell...application, our Viruses as Biological Tools blog can help you learn about the different types of viruses and pick...delivery of the actual virus to which immunity is desired. Instead, components of the virus are now delivered...What viruses can do for you Whether you are interested in basic science or clinical therapy—viruses could...there is more than one type of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided...Viruses are the SUVs of molecular biology – they can deliver materials to specific destinations, haul...out, etc. It’s how we understand what genes do! Viruses have several options to allow you to do exactly...

  15. Tips for Titering Your Lentiviral Preps

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    ...you should always titer your virus - that is, determine the amount of virus you actually have in your prep...producing virus or require precise titers. Additional considerations for titering lentivirus Volume and... prep. Taking time to properly titer your virus will not only ensure that your infection is designed in... involve infecting a target cell line with your virus and assaying for expression of a gene carried on...viral RNA can overestimate the amount of infectious virus in a prep as the measurement will include defective...reverse transcriptase will produce low levels of virus and can be used to track carryover. Functional titer...is considered a more accurate method to quantify virus as it only measures infectious viral particles. ...

  16. Quick Guide to All Things Lentivirus

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    ... oncogenic virus, or that the accessory factors in HIV positive cells could make the virus replication...consistency it’s a good idea to purify and titer your virus. Virus purification can be a tricky process. You have...incubate your cells with virus at different multiplicity of infection (MOI=ratio of virus to cells). Again, ...Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production....used were derived from the Moloney Murine leukemia virus (MoMLV) and consisted of two plasmids: the packaging...integrated into the new viral particles and the virus particles produced could not replicate. However,...Mother Nature found a way to re-create a functional virus. Indeed, by testing several vectors and packaging...

  17. Rabies and Neuronal Tracing

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    ...forms of these viruses typically used in research aren’t able to replicate, and for a virus to cross a synapse...deletion-mutant rabies virus. Retrograde neuronal tracing with a deletion-mutant rabies virus. Pubmed PMID 17179932...neuroscience. In this post, I’ll describe how rabies virus (RABV) can be used in the brain to visualize how...of cargoes, genetically engineered neurotropic viruses emerged as powerful tools for visualizing neural...connectivity (Nassi et al. 2015). The recombinant viruses typically used in neuroscience research (e.g., ... to another, it must replicate. Modified Rabies Virus: Converting a transsynaptic neuronal tracer into...and retrograde transmission occur, spreading the virus through higher and higher order neural connections...

  18. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    ... MoMLV (Moloney Murine Leukemia Virus) or MSCV (Murine Stem Cell Virus) sequences. Packaging genes (viral...range of cells a virus can infect. Note: While both lentiviruses and γ-retroviruses use the same gene...differences between engineered γ-retroviruses and lentiviruses. First, these viruses were derived from different...target cell does not produce additional virus. Are γ-retroviruses self-inactivating (SIN)? They can be....prompted the development of multiple virus packaging methods. Across these methods, the following components...envelope plasmids, such as VSV-G (Vesicular Stomatitis Virus glycoprotein), however, may be used across both ... envelope plasmids can direct the production of virus with various tropisms.  Packaging using helper-free...

  19. Top Requested Lentivirus and AAV of 2016

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    ...requested lentivirus and AAV of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9...and began delivering ready-to-use lentivirus and adeno-associated virus (AAV) to scientists around the world...expanded our viral inventory to 25 lentiviruses and 25 AAVs. These viruses have been distributed in over 200...Addgene, productivity is infectious. Curious which viruses researchers have found the most useful so far? ...cell line of choice after transduction with this virus. While various Cas9 mutants have been engineered...and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate Cas9-expressing cell lines...current AAV inventory consists almost entirely of viruses expressing DREADDs from Bryan Roth’s lab at the...

  20. An Introduction to Adenovirus

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    ... market). Although host-virus interactions likely seem important for any virus, we will see that they ...by either SARS-CoV-2, Adenovirus type 41 (AdV-41), or Adenovirus-Associated Virus type 2 (AAV2) – or some...saying that host-virus interactions are an important part of the biology of adenoviruses (AdVs). AdVs are...-stranded DNA viruses, with a linear genome between 26–45 kb in length. Human adenovirus type 5 (hAdV-...otherwise clear the virus from the cell population (Kafri et al., 1998). In fact, adenovirus gets its name ...viral hepatitis it is caused by the Hepatitis A Virus, for which a vaccine is available. However, here...Adenoviridae family, which is a very diverse family of viruses with broad host range and tissue tropism (Borkenhagen...

  21. Getting the Most from Your Lentiviral Transduction

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    ... for lentiviral infections, produce the virus, titer the virus (read this blog post for tips on titering... cell lines and virus properly and be consistent when following protocols. Need Virus? Check out Addgene's...lentiviral infection, happy virus is the second. When harvesting lentivirus, some labs prefer to collect...collected lentivirus for downstream applications. While this ensures that the highest titer virus is used...infection.     Great transduction start with great virus If having happy cells is the first step of a successful...tends to result in a higher volume of lower titer virus. In such cases you may want to consider concentrating...at -80°C. Some reports suggest rapidly freezing virus in a dry ice/ethanol bath or liquid nitrogen prior...

  22. Tips for a 1st time AAV user (by a Rookie AAV user)

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    ...concentrate the purified virus. This process can be completed in one long day, or the virus can be stored at ...Learn how Addgene produces AAV and lentivirus Check out Addgene's virus protocols Find AAV for your research...worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit intimidating... transgene of interest that’s packaged into the virus. Two to five days after transfection, the AAV-containing...have a bit extra AAV than to have to produce more virus because I don’t have enough AAV. Compared to the...precipitation, but for my experiments, I get enough virus from the cell pellet alone. If yield is important...in a 95% ethanol and dry ice bath to release the virus particles from the cells. Cell lysates are then ...

  23. Popular Retroviral Vectors and Their Uses in Scientific Research

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    ...wondering which virus fits you experiments best have a look at this viral vector overview. Viruses of the Retroviridae...Moloney Murine Leukemia Virus (MoMLV, MMLV, MuLV, or MLV) or Murine Stem Cell Virus (MSCV) genomes whereas..., for example gamma-retrovirus, lentivirus, Adenovirus, and Adeno-associated virus (AAV). If you are wondering... Viruses are intracellular parasites and natural vehicles for genetic information. Therefore they make...vectors are derived from the human immunodeficiency virus (HIV) genome. For research purposes, retroviral ...retroviral plasmids (based on the Murine Stem Cell Virus, MSCV) each with a unique 6 nucleotide barcode. ...puromycin resistance cassette. No time to produce virus? Get the ready-to-use lentiviral prep from the Addgene...

  24. Addgene’s Viral Service - Why Virus? Why Now?

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    ...Research Using Virus in Your Research - A Primer for Beginners Resources at Addgene.org Find Virus Protocols...infrastructure capabilities to produce and distribute virus. Addgene is a 501(c)3 nonprofit and our plasmid ...worked closely with us to functionally test our virus. Ed Boyden’s lab from MIT and Alla Karpova’s lab...time, dedication, advice and encouragement. Why virus? Why did we decide to take on this project? Addgene...provides viral vector technical service (details about virus production protocols and quality control can be ... developing expertise in answering questions on virus and helping provide educational resources on this...Our Viral Vector Pages Find Virus for Your Research ...

  25. Viral Vectors 101: An Introduction to AAV

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    ...Adeno-associated virus (AAV) is a single stranded, Parvoviridae DNA virus, packaged in a non-enveloped...Addgene Blog New to Virus Work? Check out Our Primer for Beginners Read Our Lentivirus FAQ Learn about Viral...package an rAAV vector; these capsids allow the virus to infect (or not infect) different cell types with... heftier genes, you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb. Co-infection...AAV data hub to see if others have used relevant viruses for your system. This post was contributed by...al. "An essential receptor for adeno-associated virus infection."Nature 4:520(7588):108-12 (2016). PubMed...Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently...

  26. 10 Great Guest Posts We're Thankful For

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    ... tool you can use to study basic virus biology while leaving virulent components behind. Minigenomes are...function. Adapting Toehold Switches to Detect Zika Virus The RNA-based toehold switches described in this...engineered to bind RNA sequences derived from Zika virus and, upon binding, produce reporter proteins. These... be used to quickly detect the presence of Zika virus. Read this post from Keith Pardee and Alexander ...amazing tools may be used to rapidly diagnose Zika virus infection. Microbiology and virology Minigenomes...Minigenomes - A Safe Way to Study Dangerous Viruses Contributed by guest blogger Tessa Cressy from the labs of...for Gene Expression in Mammals Adeno-associated viruses (AAVs) are commonly used to deliver a genes of ...

  27. AAVs CREATed for Gene Delivery to the CNS and PNS

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    ...inducer virus, but the desired sparsity of labeling is achieved with lose doses of inducer virus. Additionally...pain. While many new capsids (i.e. the part of the virus that determines tropism) have been developed that...sites flank the downstream polyA sequence. When viruses produced from this plasmid pool are injected into...into the bloodstream allows for dispersion of the virus but it has to be able to cross the blood-brain barrier...green neurons near the arrows. When lower doses of virus are used in order to decrease the number of neurons...either an AAV-PHP.B or an AAV.PHP.eB capsid. Both viruses were IV injected at a dose of 1x1011 vg per mouse...into either an AAV.9 or an AAV.PHP.S capsid. Both viruses were IV injected at a dose of 1x1012 vg per mouse...

  28. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

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    ...Your Lentivirus Preps Additional Resources on Addgene.org Check out the Viral Service Find Virus Protocols...wondered whether or not the dry ice we use to keep viruses frozen during shipment was having an impact on ...dry ice, and specifically how dry ice can affect virus from Addgene. Bottom line: there’s good news and...storage container for 48 hours (to replicate how our viruses are shipped), where they would be exposed to a ... to pH 5.1 did not alter the infectivity of the virus. However, there was a substantial reduction in infectivity...seconds prior to thaw, and also using the thawed virus within 24 hours. References 1. Murphy BM, Swarts...purification protocol for recombinant adeno-associated virus vectors. Mol Ther Methods Clin Dev. 2014 Aug 13;...

  29. Congratulations, Deck The Lab winners!

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    ...decorated with dozens of individual virus snowflakes, since snowflakes and viruses both exhibit similar forms ...Winter Wonderlab🎶 We decided to #DeckTheLab with virus snowflakes from @CVRHutchinson #SciArt #STEAM #WinterWonderlab...displaying over 25 archaea, bacteria, fungi, and viruses. Final tree with the complete #MicrobeAdventCalendar...plus more!) It features archaea, bacteria, fungi, viruses, and one microbe ornament even dyed with real microbial...

  30. A Scientist's Guide to Ebola

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    ...not the virus itself that kills you, but the cytokine storm it ultimately sets off.  The virus works its...negative-sense, single-stranded RNA genomes. The virus does all of its damage with just seven structural...evidence in PLoS Pathogens shows that the Ebola virus glycoprotein remains essential in allowing those...within host cells and turn the cell into a veritable virus factory.  Their findings were reported in the Journal...resources. Ebola Basics A few basics: Ebolavirus (EBOV) is a Filovirus, a family characterized by single,...

  31. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    ... the reprogramming factors Sendai virus- a non-integrating RNA virus Episomal vectors- extrachromosomally...laboratories use Sendai virus to reprogram PBMCs efficiently [5]. However, the Sendai virus method is expensive... of virus-free hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai virus requires...reactions. Another drawback of using the Sendai virus is the persistence of residual viral material [6...concerns involved in production and usage of Sendai virus. Reprogramming efficiency of PBMCs with episomal...provides a practical way to generate transgene-free, virus-free clinical grade hiPSCs from easily accessible...using a combination of activated T cells and Sendai virus. Nat Protoc, 2012. 7(4): p. 718-28. PubMed PMID:...

  32. Viral Production at Addgene

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    ...   Viruses are generated using standard methods that have been optimized for each specific virus in order...experiment using our virus protocols. Browse our AAV vector collection. Lentivirus Production Lentiviral...high quality preparations. After production, all virus preps are titered and subjected to quality control...preps available from Addgene. After receiving your virus from Addgene, read our detailed recipient instructions...contamination. In the event of contamination, all of the virus produced in the line will be taken offline and discarded... viral production. Barcoding allows our DNA and virus to be monitored throughout production, ensuring ...available from Addgene. After receiving your virus from Addgene, read our detailed recipient instructions for ...

  33. The Importance of High Titer for AAV Transductions

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    ... comes to infecting cells, titer, the amount of virus used, really does matter. (*psst*, quality definitely...essentially nonexistent, as there wasn’t enough virus to produce any observable expression. Doubling this...address the issue of low titer by concentrating your virus. See our Concentrating AAV Protocol (Step 10) for...instructions on how to do this. Concentrating your virus will allow you to add many more viral particles ...Recent advances in recombinant adeno-associated virus vector production." Kidney international 61.1 (2002...BR, Chamberlain JS. Recombinant Adeno-associated Virus Transduction and Integration. Molecular therapy ...-Use AAV Preps Find Lab Protocols for Work with Virus Watch Our AAV Purification Video ...

  34. With Our New Viral Service, We're Taking CRISPR Further!

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    ...provide CRISPR as virus? “Like all things in science, getting a high titer Cas9 lentivirus prep takes a bit... only the starting point for the production of viruses used to deliver CRISPR components to cells or organisms...study. In the past we’ve left the arduous task of virus production to individual labs, but now we’re very...pathways. Find Validated gRNAs to use with your Cas9 Virus Pooled libraries now available in lentiviral format...discoveries." If you ever run out of ready-to-use virus and want to make new lentiviral preps on your own...to distribute their CRISPR tools as ready-to-use virus and, in particular, the Church lab at the Wyss Institute...the Most from Your Lentiviral Transdution Using Virus in Your Research: A Primer for Beginners 5 Tips ...

  35. Viral Vectors 101: The AAV Retrograde Serotype

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    ...soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected into the brain and...Ready-to-use Virus for Your Research Visit Our Lentivirus Guide Page ...tools, making them useful tracers, but finding a virus that works well for retrograde tracing can be difficult...difficult. The in vivo workhorse adeno associated virus (AAV) shows relatively low retrograde transport ...transport efficiency in most serotypes. Some viruses (e.g., rabies) naturally exhibit retrograde transport and... virus...Addgene Blog Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral Service ...

  36. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

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    ...response as the target of vaccines and allows the virus to gain entry into human cells via human ACE2 receptors...This result was similar to studies that used live virus (Cele et al., 2021). “We weren't completely shocked...these results] because we were simulating the live virus experiment,” says Balazs. “But what was surprising...SARS-CoV. These two viruses are quite genetically distinct from SARS-CoV-2 and its variants. Yet, the B...outcomes of convalescent sera with various pseudotyped virus. Image from Garcia-Beltran et al., 2021 with permission... sufficient after one vaccine to neutralize the virus,” says Balazs, emphasizing the importance of the... COVID-19 collection Find SARS-COV-2 pseudotype virus plasmids from Addgene ...

  37. Overcoming the Challenges of Lentiviral Production

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    ... not just the lentivirus; consequently, this may not be the best method when pure virus is required downstream... best fit for the experiment, trying to produce virus of a usable titer, or fine-tuning selection and ...packaging cell line and thereby affect the titer of the virus. Additional plasmids will also affect experiment...harvest approach at 48 and 72h produces high titer virus for transfer plasmids with small inserts (such as...-fold meaning that a 10 cm2 dish preparation of virus will only yield 150 μl of material as opposed to... if the experiment requires a certain volume of virus, consider scaling up by either increasing the number...experiment. Media choice, reaction vessel, dose of virus, volume and timing of transduction will all impact...

  38. All in a Twist: dsRNA

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    ...within the host. dsRNA viruses are inherently double stranded, while ssRNA viruses can generate dsRNA intermediates... store their genetic information. All of these viruses have one thing in common – they can generate dsRNA...as part of the viral replication process. dsDNA viruses yield dsRNA through a slightly different mechanism...that of vaccines. If you know the sequence of a virus, you could introduce some of its dsRNA intermediates... response and then, hopefully, immunity to that virus. This system is certainly a double-edged sword –...Untwisting RNA is important for therapeutic delivery, viruses, human disease, and much more! Resources and References...

  39. Delivery Methods for Generating iPSCs

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    ...Sendai viral vectors: Sendai virus is a single stranded, negative sense RNA virus. It’s a member of the Paramyxoviridae...generated with Moloney murine leukemia virus (MMLV)-derived retroviruses. Retroviral vectors, such as pMXs ... No genomic integration Inefficient Sendai Virus Non-integrating Medium Medium Generates transgene-free...genomic integration Can be difficult to fully remove virus from cells Plasmid Non-integrating, usually Medium.... This can be overcome by using a Tet-inducible virus to allow for controlled expression of reprogramming...Paramyxoviridae family of viruses, which also includes measles and mumps. Sendai transduces a wide range...replication competent, it’s difficult to eliminate the virus from all cells, even after many passages. Ban et...

  40. Help Us Name Blugene’s New Virus Friend

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    ...hashtag #BlugeneVirusFriend on Twitter. As with Blugene’s name, we’d like to give their virus friend a... your help in finding a good name for Blugene’s virus friend.  During the month of January, we’ll be...swag. Have fun! Suggest a name for Blugene's new virus friend here Additional resources on the Addgene...

  41. Viral Vectors 101: Producing Your rAAV

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    ...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...

  42. Adeno Associated Virus (AAV) for Cell and Gene Therapy

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    ... 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...

  43. Scientists Map the SARS-CoV-2-Human Interaction Network

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    ...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...

  44. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...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...

  45. Meet Aavery, Blugene's New Virus Friend!

    Type
    Blog Post
    ...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...

  46. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...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...

  47. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...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...

  48. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...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...

  49. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...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 ...

  50. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...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...
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