... 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...

...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 ...

...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...

...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 ...

...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...

... 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 ...

...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...

...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;...

...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...

...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,...

... 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:...

...   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 ...

...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 ...

...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 ...

... 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 ...

...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 ...

... 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...

...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...

...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...

...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...