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Showing: 51 - 100 of 319 results

  1. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    Blog Post
    ... 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 (...

  2. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...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...

  3. Adenoviral Vector Production and Troubleshooting

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    Blog Post
    ...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...

  4. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...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...

  5. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...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...

  6. Viral Vectors 101: Parts of the AAV Packaging Plasmid

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    Blog Post
    ...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...

  7. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

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    Blog Post
    ... 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...

  8. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...= 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 ...

  9. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ...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...

  10. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...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 ...

  11. Viral Vectors 101: AAV Variables That Matter

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    Blog Post
    ... to just dive right in and start injecting your virus, a successful AAV experiment starts with validation...compositions of AAV capsids, which determine how a virus enters the cell. Different serotypes utilize a number... serotypes have broad tropism (the ability of a virus to enter a particular cell type), but there are ...begin testing, you’ll next need to decide how much virus to put into your system, which is controlled through...picked, since that can affect the spread of the virus.  A better way to define high titer, then, is this...recombinase-dependent vectors) increases with the amount of virus you add, so it is strongly recommended to optimize...(2018). An Alternate Route for Adeno-associated Virus (AAV) Entry Independent of AAV Receptor. Journal...

  12. Important Considerations When Using AAVs

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    ...mutagenesis in humans (1) and AAVs are non-enveloped viruses, so they are resistant to alcohol-based disinfectants...freezing and thawing. Although more stable than other viruses, multiple freeze thaw cycles can still reduce AAV... structure (scAAV) (7). As single stranded DNA viruses, AAVs normally require second strand synthesis ...phenotype. Previous neural tracing methods used viruses that were potentially toxic and would result in...4. Castle, Michael J., et al. "Adeno-associated virus serotypes 1, 8, and 9 share conserved mechanisms...Self-complementary recombinant adeno-associated virus (scAAV) vectors promote efficient transduction independently...efficient transduction by recombinant adeno-associated virus vectors." Journal of virology 70.5 (1996): 3227-...

  13. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    Blog Post
    ...poorly understood,” says Qiu.   Qiu developed a dual virus system using both optogenetics and chemogenetics...neurons is in progress, she has validated the dual virus method to study CR neurons which are depicted here...Schematic diagram of the construction of the dual virus and the experimental approach. (B) CR-Cre mice demonstrate... response due to blue light stimulation of dual virus-expressing CR neurons. (C) Duration of painful responses...) Immunohistochemistry verification of the dual virus expression in CR neurons of the spinal cord. Created...

  14. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    ...plasmid which contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains...Cre-Lox system.  WPRE The Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE...Bachle SM, Kamens J, Fan M (2019) Adeno-Associated Virus Technologies and Methods for Targeted Neuronal Manipulation...the Inverted Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods 30:206–213 . https:/...

  15. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    ...engineering viruses Acr genes can be used as positive selectable markers in engineering viruses. For example...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles...Papayannopoulou T, Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases...

  16. Viral Vectors 101: Inverted Terminal Repeats

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    ...Adeno-associated viruses (AAV) are an important biological tool that allow scientists to express a gene...single stranded DNA genome of AAV in the native virus. In an AAV vector plasmid, they flank the gene of... L., & Samulski, R. J. (2020). Adeno-Associated Virus Serotype-Specific Inverted Terminal Repeat Sequence...the Inverted Terminal Repeats of Adeno-Associated Virus. Hum Gene Ther Methods, 30(6), 206–213. https://...Repeat Integrity on rAAV8 Production Using the Baculovirus/Sf9 Cells System. Hum Gene Ther Methods, 28(5...

  17. Viral Vectors

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    Collection
    ...generate virus (right), similar to how natural viruses can be delivered to a host and generate virus (left...more viruses. When a virus infects a host cell, its genome directs the production of more viruses. However...Type of Virus Should I Use? Use this guide to learn more about the different types of viruses that are...genome. Adeno-associated Virus (AAV) Plasmids | Guide Adeno-associated viruses (AAVs) are commonly used... most popular virus for gene transfer in clinical trials. These viruses have a relatively high transduction... DNA for use with virus-mediated delivery has many advantages for research. Viruses facilitate the delivery... they give rise to viruses with modified viral genomes. Naturally occurring viruses harbor genetic material...

  18. Lentivirus ddPCR Titration

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    Protocol
    ...Transducing Cells Thaw the virus on ice. Prepare the following dilution series of virus in DMEM complete: Prepare...the first virus dilution, add 160 µL of viral stock to 240 µL of media. When adding the virus to the diluent...the following virus dilutions were used to transduce cells: Initial dilution factor of virus stock Dilution...titer, TU/mL V = Viruses per genome C = # Cells per well D = Dilution factor v = Virus volume, mL *In the...Titration by qPCR Addgene Viral Service Lentivirus Guide Virus Blog Posts Introduction This protocol describes...diluent, pipette 10 times, to remove virus from pipette tip. To mix the dilution, set the P200 pipette...as indicated in the chart below. Dilution Factor Virus Volume to Dilute Media Volume Total Volume 2.5 160...

  20. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

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    Protocol
    ...analyzing your plasmids. Virus Protocols for titering and testing your virus preparations. Antibodies ...protocol Fluorescence Titering Assay for Lentivirus Quantify virus using fluorescence measurements Colony...Colony Formation Titering Assay for Lentivirus Quantify virus by counting antibiotic resistant colonies Generating...physical titer of your adeno-associated virus preparation Lentivirus Titration by ddPCR Use ddPCR to measure...consortium for targeting human and mouse genes. Virus Name Description Link to Video General Transfection...Production in HEK293 Cells Produce adeno-associated virus to deliver your gene of to the target of your choice...Gradient Ultracentrifugation Purify adeno-associated virus from a preparation produced in mammalian cells Watch...

  22. AAV for Neuronal Tracing

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    ...for rabies virus-based monosynaptic tracing. These AAV can be used with modified rabies virus to perform...that lack the G protein, the virus can replicate but the newly formed viruses are unable to infect any neighboring...envelope protein of (EnvA) of leukosis virus. EnvA can direct virus infection specifically into cells that...neuronal tracing, to be used with modified rabies virus. Viral...tracing. Note : Addgene does not distribute rabies virus. AAV Vectors for Monosynaptic Neuronal Tracing ID...Can be used to complement deletion-mutant rabies virus AAV1 Wickersham 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA...can be used to complement deletion-mutant rabies virus AAV1 Wickersham 100799 pAAV-TREtight-mTagBFP2-B19G...

  24. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ...advantages of viruses and naked DNA while eliminating some of the drawbacks. Specifically, viruses are able...certain forms of gene therapy - avoiding the use of viruses is also important for social and regulatory reasons...

  25. Addgene Protocols Go Viral: AAV Purification and Titration

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    ... serve the scientific community. Here are a few virus-related protocol videos that may help to save you...produce vital tools in the lab such as antibodies or viruses. If you’re new to tissue culture, check out this...

  26. Finding nucleic acids with SHERLOCK and DETECTR

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    ...interest, for example, a sequence specific to a virus or pathogen. Once Cas13 recognizes and binds to ... distinguish between Zika and a closely-related virus, Dengue from multiple sample sources. SHERLOCK could...including clinical diagnostics (e.g. pathogen or virus detection), therapeutics and sensitive genotyping...specific DNA sequence, such as the Human papilloma virus (HPV) genome, via a crRNA. An ssDNA-fluorescently...Learn about using toehold switches to detect Zika virus   Learn more about targeting RNA with Cas13a (C2c2...

  27. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

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    ... human lung cancer cell line), armed only with viruses obtained through Addgene’s viral service and the...repeatability and bad news for a screen. Because Addgene viruses are provided at a specific titer, you can better...expression was the most efficient for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011...activity in my stable A549 cell pools. Using this virus, it’s expected that Cas9 activity, guided by the... I just randomly chose one of my stable pools. Viruses containing BRAF gRNA (BRDN0000561167 and BRDN0001146266...regions of your gene. I used existing BRAF gRNA viruses, but you can find a list of validated gRNA plasmids...Lines with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus Production...

  28. AAV Vector Quality Control: Going the Extra Mile with NGS

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    ...the purification step because they are inside the virus itself. While these impurities are generally considered...Prokaryotic Sequences during Recombinant Adeno-Associated Virus Production and Their in Vivo Persistence after Vector... in rAAV Vector Preparations by Single-stranded Virus Next-generation Sequencing. Molecular Therapy - ...Control Testing of Recombinant Adeno-Associated Viruses. In: Methods in Molecular Biology. Humana Press....org Find Ready-to-Use AAV Find Ready-to-Use Lentivirus Browse Viral Plasmids ...

  29. Colony Formation Titering Assay

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    Protocol
    ...Formation Titering Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols...the seeding of the cells at the same time as the virus-mediated transduction. For some cell lines, transduction...into 20 mL of media. If using freshly collected virus, filter through a 0.45 μm polyethersulfone filter...freeze-thaw. If you are freezing and aliquoting virus, it is recommended that you titer from the frozen...titer associated with freeze-thaw. If using frozen virus, thaw the lentiviral aliquot on ice prior to use...Complete Containing 10 μg/mL Polybrene (μL) Volume of Virus Added to Plate (μL) Volume of Cells Added to Plate...Plate (μL) Final Viral Dilution 1:10 100 of Stock Virus 900 150 1,350 1:100 1:100 100 of 1:10 900 150 1,...

  31. History of CRISPR Cas - A tale of survival and evolution

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    ...adaptive defense strategy against life-threatening viruses. Evolving to survive Although the CRISPR sequences...ticket to ensure prokaryotic survival; however, viruses, too, are famous for their ability to devise new...strategies to reproduce within a host cell. A few virus particles have learned to circumvent the CRISPR/...

  32. Lentiviral Vector Uses and Overview

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    Blog Post
    ...viral vector systems based on retroviruses like Moloney murine leukemia virus (MMLV). The vectors could integrate...Resources on Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors...possibility of creating a replication-competent virus via recombination between the transfer and packaging...events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition ...useful viral vectors in the lab. Advantages of lentivirus include a large genetic capacity and the ability...dividing cells. Another type of vector, based on adenovirus, could infect non-dividing cells, but without... Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to infect non-dividing cells...

  33. Fluorescence Titering Assay

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    Protocol
    ...cells with fluorescence V V = Virus Volume, mL For example: If 15 µL of virus added to 150,000 cells resulted...Fluorescence Titering Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols...the cells overnight. If using freshly collected virus, filter through a 0.45 μm polyethersulfone filter...freeze-thaw. If you are freezing and aliquoting virus, it is recommended that you titer from the frozen...titer associated with freeze-thaw. If using frozen virus, rapidly thaw the lentiviral aliquot on ice prior...the dilution factors (method 1) or the volume of virus (method 2): Method 1 Method 2 $$T = {N*F*D\over ...used in downstream applications. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please...

  34. SARS-CoV-2 Pseudotyped Virus

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    Collection
    ...components of one virus (such as HIV, murine leukemia virus, and vesicular stomatitis virus G) are packaged...the study of the live virus. Alternatively, it is possible to generate pseudovirus, where the structural...suitable for generating pseudotyped virus for SARS-CoV-2 and other coronaviruses using appropriate packaging ... Protein Nemazee lab - Pseudotyped virus protocol for coronaviruses (184 KB) Addgene Blog Posts Viral ...-19 - Viral Pseudotyping SARS-CoV-2 Pseudotyped Virus Mechanisms of SARS-CoV-2 infection remain poorly...packaged as replication-defective virus particles with the SARS-CoV-2 spike (S) glycoprotein on the envelope...envelope surface. Pseudotyped virus particles can then be used to infect permissive cells that express ACE2...

  35. Adenovirus Plasmids

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    ...Plasmids Additional Resources: Adenovirus Guide Virus Biosafety Lentiviral Plasmids AAV Plasmids Viral Vectors...101 eBook Adenoviruses are a class of medium-sized, non-enveloped, double-stranded DNA viruses known to...levels of transgene expression. Additionally, these viruses are able to target a broad range of dividing and... Bunz 16402 pShuttle Shuttle For production of viruses containing transgene under a chosen promoter Vogelstein...Vogelstein 16403 pShuttle-CMV Shuttle For production of viruses containing transgene under CMV promoter Vogelstein...pAdTrack Shuttle For production of GFP-trackable viruses containing transgene under a chosen promoter Vogelstein...pAdTrack-CMV Shuttle For production of GFP-trackable viruses containing transgene under CMV promoter Vogelstein...

  36. AAV Purification by Iodixanol Gradient Ultracentrifugation

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    Protocol
    ...ultracentrifugation to get pure adeno-associated virus (AAV) preps for use in your research....Ultracentrifugation You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction This protocol...an isomolar density gradient medium suitable for virus purification and isolation of cells, organelles,... remove iodixanol prior to use of your purified virus. Nonetheless, we recommend performing a buffer exchange...fewer fractions once you have a good idea when the virus elutes from the gradients. In a biosafety cabinet..., or you will lose a significant amount of your virus. When the first fraction is collected, move the ... off the walls of the filter to recover as much virus as possible. Store at 4 °C for short term (2 weeks...

  37. Newly Updated AAV Data Hub!

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    Blog Post
    ...labelled with an AAV1 Cre virus combined with a Cre-dependent AAV9 tdTomato virus in the mPFC.  The AAV9 ...

  38. Virus Protocol - Generating Stable Cell Lines

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    Protocol
    ...they vastly outnumber the virus particles. Incubate the cells with the virus for 48–72 h. Gently aspirate...Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral Service Introduction This protocol can be...downstream applications. If you’ve titered your virus beforehand, you can narrow this range according ...6-well plate will hold 5 dilutions plus one 'no virus' control well). Perform a "reverse transduction"...added to the wells that already contain 0.5 mL of virus solutions at various dilution. Make sure to use ...Transducing too many cells relative to the number of virus particles reduces the transduction efficiency, resulting...Typically, cells transduced with lower dilutions of the virus will have higher levels of expression. Consider ...

  39. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    ...brain, researchers inject genetically modified viruses encoding microbial rhodopsins into specific cerebral...and Maguire, 2016). Recombinant adeno-associated viruses (rAAVs) have become increasingly popular for gene...cells? This can be achieved using Cre-dependent viruses, such as the optogenetic FLEx vectors in combination...cell-type promoter. Once injected into the brain, the viruses infect all the cells with an inactive opsin gene...References Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary...

  40. Download Science-Themed Zoom Backgrounds from Addgene

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    ...working with AAV or lentivirus in your research, you might want to use one of our virus backgrounds. Addgene...barcoding libraries, and more. Download the green virus background.           Maybe your viral research...well as transfer plasmids. Download the orange lentivirus background.         We also have ready-to-use...background for you, too! And we have a COVID-19 and coronavirus collection page, where you can find plasmid tools...

  41. 15 Years of Addgene: The Top 15 Plasmids

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    Blog Post
    ...which types of cells a virus can infect. The VSV-G envelope protein enables this virus to infect many different...lentiCRISPR v2 great? It produces a ~10 fold higher titer virus, making it ideal for CRISPR pooled library screening...posts Read our viral vector blog posts Read our lentivirus guide Resources on the Addgene website Browse...

  42. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    Blog Post
    ...introduced into bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages ...can be done with lentiviral and Adeno Associated Viruses (AAV). Lentiviral and AAV can be used to create... basic molecular biology, plasmid cloning, and viruses. References Baltrus, David A., and Karen Guillemin...bacteria have gained antibiotic resistance and virulence. Scientists have studied how HGT occurs in nature...

  43. Clear and Accessible Writing

    Type
    Blog Post
    ...ultracentrifugation so that a scientist can extract virus from the correct layer.  Third, think about the ...at the interface of the 40% layer containing the virus and 60% iodixanol layer, with the bevel pointed ... the tube allows air to enter the column so the virus will flow out the bottom syringe."Feel free to share...

  44. A Primer on Optogenetics: Introduction and Opsin Delivery

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    Blog Post
    ...that offer his constructs. Importantly, control viruses (not expressing the opsin) are also available. ... optogenetics experiment you may need to inject virus into 20 or more mice and the injections must be ... practice with die injection prior to using the virus. You may need to adjust coordinates based on your...co-localization immunofluorescence, cell-type specificity. Viruses that only express the fluorescent proteins are ...affects the length of time you may need to allow the virus to express. For example, if you inject into the ...

  45. Adeno-associated virus (AAV) Plasmids

    Type
    Collection
    ...Adeno-associated viruses (AAV) are small viruses originally discovered as contaminants of adenovirus stocks. One... presence of another "helper" virus, such as adenovirus or herpes virus, in order to propagate. This is...Viral Plasmids Adeno-associated Virus (AAV) Plasmids Adeno-associated Virus (AAV) Plasmids Additional Resources... requirement has been circumvented with “helper-virus free systems,” which enable the production of infectious...infectious AAV particles without the use of a helper virus. Instead, specific gene products can be provided...infectious AAV particles, a helper plasmid (encoding adenovirus E4, E2A and VA) and a RepCap plasmid (encoding... Helper plasmid for AAV packaging, expressing adenovirus E4, E2A and VA James M. Wilson RepCap Plasmids...

  46. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies such that the virus is undetectable... undetectable in the blood. But the virus doesn’t completely disappear; it just hides in latently infected...infection, which would make it easier for an “escape” virus to propagate and infect adjacent cells. It will ...of viral DNA efficiently suppresses hepatitis B virus. Sci Rep. 2015 Jun 2;5:10833. PubMed PMID: 26035283...

  47. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...delivery vehicle that’s based on mouse leukemia virus (MLV) virus-like particles (VLPs). VLPs lack viral genomes...box.  Nanoblades are engineered murine leukemia virus (MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins...with a mix of VSV-G and the baboon endogenous retrovirus Rless glycoprotein (BaEVRless) had high rates...

  48. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...of the upstream efficiency element of the simian virus 40 late polyadenylation signal by using in vitro...Donello JE, Trono D, Hope TJ. Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element Enhances ...H-G. Multiple copies of the Mason–Pfizer monkey virus constitutive RNA transport element lead to enhanced... Substantially Increases Expression Levels of Lentivirus-Delivered Transgenes but Has the Potential to...

  49. Lentivirus Production

    Type
    Protocol
    ...replace with fresh media. Day 3-4 (am): Harvest virus Equipment Class II, Type A2 Biological Safety Cabinet...DMEM Complete or OptiPro SFM. Incubate the cells. Virus can be harvested at 48, 72, and 96 h post transfection... Protocols Lentivirus Production Lentivirus Production You may also ...this protocol to generate lentivirus Protocols...Introduction This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T...lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications...

  50. Microbiology Resources

    Type
    Collection
    ...Plasmids for Viruses Species Dengue Hepatitis Influenza Mumps virus Rubeola (measles) West Nile virus (WNV) ... your microbe of interest, including bacteria, viruses, protozoa, fungi, and more. Plasmid...range of microbial fields, including bacteria, viruses, parasites such as protozoa, and fungi. Find plasmids...complementation constructs - Lahaye Lab Vaccinia virus ORF library : Expression library that contains each... the predicted open reading frames for vaccinia virus - Bennink and Yewdell Labs Yarrowia lipolytica Golden...
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