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... download the PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging ...corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier to plan...for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting...right, you need to do a few calculations. This calculator can do the math for you and it’s simple to make...whole experiment And here’s the equation the calculator uses to give you the μl needed of each plasmid...experiments. Let us know what you think about the calculator in the comments section below. References 1....Production of Recombinant Adeno‐Associated Viral Vectors for In Vitro and In Vivo Use." Current Protocols...

...Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors ... Viruses are essentially DNA (or RNA) delivery vectors, so what makes them better than the alternatives...measures in a separate, upcoming post. If a viral vector seems like the right approach for your experiment...experiment?           Click Here to Find Viral Vector Resources at Addgene! References and Resources...S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed... J, et al. “Altering the tropism of lentiviral vectors through pseudotyping.” Curr. Gene. Ther. 5 (2005...

...Science History Rap Battle A great example of science story telling following the unfortunate story of Rosalind...and their importance in understanding biological history. 4. Please Don't Kill My Hive Beautifully filmed...with us. Tom McFadden is a YouTuber and science educator looking for creative ways to implement the next...

...When our Director of Biology, Lianna Swanson, suggested that I might be able to attend the 10 Years of...Oct4, Sox2, Klf4, and cMyc (the so called OSKM factors). As the Yamanaka lab's primary Addgene contact... forward to seeing how the sharing of his OSKM factors have affected the stem cell community – a community...the first clinical trial with iPSCs with his collaborator Takahashi in 2014 – just 8 years after the OSKM...control process alone takes nearly a year. Addgene depositor Juan Carlos Izpsua Belmonte said it took the joint...effort of five post-docs to implant human kidney progenitors into a pig. In stark contrast to recent genome...the scientists I've already mentioned, Addgene depositors Marius Wernig, Kristin Baldwin, Kathrin Plath...

...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12 ...addition to the combinatorial expression of other genetic elements including transcription factors (Cheng et...indicated below and pool-clone them into the pAWp28 storage vector (Figure 1): Forward oligo: 5’- CACCGNNNNNNNNNNNNNNNNNNNGTTTGGGTCTTCGAGAAGACCTATTCXXXXXXXXC...barcode, respectively. Once cloned into the storage vector, the pooled, barcoded gRNA library is now ready... an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your favorite biological...screening are vastly useful for mapping the combinatorial genetics that underlie complex regulation in...barcoded gRNA libraries that can be used to combinatorially modify the genome, screen for a particular ...

...Initial experiments exposed S. thermophilus to predatory phages to test if the exogenous DNA would be incorporated...system that utilized various cas genes not only to store a record of invading phages but to also destroy ...race" between phages and prokaryotes; the ultimate victor will be the species who has the strongest will ...

... growth factor receptor 1 (FGFR1)3 αVβ5 and α5β1 integrins3 Hepatocyte growth factor receptor (HGFR)3 ...3 AAV receptor (AAVR)2 Epidermal growth factor receptor (EGFR)3 AAV8 Laminin receptor (LR)3 ... Table 1: AAV capsids and receptors. Capsid Primary receptor Coreceptor AAV1 α2,3-/α2,6 N-linked...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...serotypes have a primary cellular receptor and sometimes one or multiple coreceptors, which may be present on ...acids1 AAV receptor (AAVR)2   AAV2 Heparan sulfate proteoglycan (HSPG)3 AAV receptor (AAVR; not ...3 Laminin receptor (LR)3 CD93 AAV5 2,3-N-linked sialic acid4 AAV receptor (AAVR)2 Platelet-derived...

...Addgene blog Viral Vector Elements The Retroviral Life Cycle Popular Retroviral Vectors and Their Use in...  Frequently asked questions about γ-retroviral vectors How do engineered γ-retroviruses differ from lentiviruses...Woodchuck hepatitis virus post‐transcriptional regulatory element; sequence that stimulates the expression...was originally published in 2018, in the Viral Vectors 101 eBook, 1st edition.  References and resources...

...been finding and sharing stories unique to our repository and resources. As editor of Addgene’s eBooks — ...amount of vectors available and number of requests tell only part of the story. Viral vectors are, after...GCaMP-encoding viral vectors available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene...information from requestors and depositors, and identifying trends in our own repository data, giving us...released Viral Vectors 101 2nd edition — I was incredibly excited when we picked our viral vector service as... as one of those stories. Viral vectors are viruses engineered to be safely used to deliver a genetic ...titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both...

...proteins from the AAV2 serotype are used to package vector genomes flanked by AAV2 ITRs into a capsid other...Strohl WR (2017) Adeno-Associated Virus (AAV) as a Vector for Gene Therapy. BioDrugs 31:317–334 . https:/...for Targeting and Improving the Efficacy of AAV Vectors. Molecular Therapy - Methods & Clinical Development...Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction. Human Gene Therapy...

...These are stormy times. It feels like we’re being buffeted by wind and waves and sometimes wonder if ...would be fully open and available to industry requestors as well. This is a time when we need as much ...materials via Addgene. Perhaps if each of many grantors supporting research could provide Addgene with...Initiative was able to step in to be our “port in the storm.”  "Addgene is a unique resource for scientists ...research is accelerating essential research in laboratories all around the world." We thank the Chan Zuckerberg...whatever lies ahead this year. We can see signs of the storm passing. We’ve recently piloted Addgene’s new cloning...cloning grade DNA service, and viral vector distribution is recovering from the pandemic-related slowdown...

...plasmid. Parts shown in grey are optional cis-regulatory elements.   Key elements of an AAV plasmid...outside of the ITRs does not get packaged in the AAV vector. The cloning capacity between the ITRs is ~4.4 ...plasmid harbors non-coding sequences. Other cis-regulatory elements Next let’s talk about some of the elements...Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when... the Addgene blog Learn more about basic viral vector elements Read about four considerations for the...

...we introduce you to the Journal of Emerging Investigators, an open-access journal that enables high school... the interview with the Journal of Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction...Olivia Ho-Shing, and the Journal of Emerging Investigators. Chapter 2 - Getting high school students and... building curricula and developing one-on-one mentorship programs. Chapter 6 and Outro - Interview with...

...welcome Julie Cicalese to the Addgene Board of Directors. Cicalese served as the Chief Human Resource Officer...School. The addition of Cicalese to the Board of Directors brings human resources experience and insight ...included the Belmont Human Rights Commission and mediator for the Massachusetts small claims court summary... Meet the other members of Addgene's Board of Directors Find careers at Addgene ...

...particles, particularly Resistance or R-factors. Like F-factors, R-factors could be transferred between bacteria...noted that, unlike F-factors, the evidence did not support the idea that R-factors could integrate into...cloning vectors such as pBR322, pACYC, and pUC were developed to provide higher copy number vectors that ...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...were working to understand genetic cytoplasmic factors that could be transferred between cells. At the...and the labels applied to them were vague or contradictory, owing in part to the fact that very little ...little was known about the role these factors played within an organism.  So how did plasmids get their name...

... bank account, business cards, incorporation, laboratory lease, etc. I still flip to that page in my lab...GMO food plants. When a press piece ran on the detector plants, we expected to see that same anger. Instead...business. We ended up applying to a business accelerator freely stating that we had an idea, knew how ...– $30,000 for research and another $30,000 in mentorship and business training. The money is a drop in...rough day, and we have great partners in this accelerator program who want us to succeed. We’re excited...excited to see what the next few months have in store for us. If you’re thinking about making the switch from...duty to pursue a degree in a synthetic biology laboratory and received her M.S. from CSU in 2014. Outside...

...during storage over prolonged periods of time, especially at room temperature or at 4°C. Storing DNA: temperature...way to store and elute the DNA. References Murakami M (2013) Evaluation of DNA Plasmid Storage Conditions...Figure 1: So many choices! What to do? The main factors that can affect DNA stability in solution are pH...and it will impact the time you will be able to store your sample without having substantial degradation...This is because divalent cations are essential cofactors for many tools used in modern molecular biology...things when you elute your DNA: where you plan to store it and for how long. The most common solution is...

...minimize it. Overview of DIO/FLEx vector design Recombinase-dependent vectors utilize pairs of specific sequences...in the expression of receptors, post-translational modifications, and other factors that AAV utilizes to...in optimizing DIO/FLEx vectors is to know the recombination rate of the vector you are using. Contact ...supplier of your vector, to obtain this information.  Optimize titer for the DIO/FLEx vector Once you have...Conclusion Using AAV vectors in your experiments requires a full understanding of vector performance in your...that there are many known and unknown factors that affect vector performance in each and every system ...recombinase-dependent AAV vectors and mitigation with cross-over insensitive ATG-out vectors. Proc Natl Acad Sci...

...blog Check out our Viral Vectors Featured Topic Page Subscribe to Viral Vectors Blog Posts Learn How We... of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous...To make sure that adeno-associated viral (AAV) vectors available at Addgene are suitable for in vivo studies...should you care about viral quality control? AAV vectors are produced by scientists - not robots. Unfortunately...are especially important when producing multiple vectors in parallel because the risk of accidental errors... or the cell genome can be packaged inside the vectors during production. These impurities cannot be removed...generally considered innocuous for research-grade vectors, we’d like to make sure they are present at a very...

...cytosine base editors (CBEs) and base changes from A•T-to-G•C are mediated by adenine base editors (ABEs). ...David Liu’s lab created the first base editor in 2016 (Komor et al., 2016) and since then has been trying...expand their precision editing capabilities. Base editors make specific DNA base changes and consist of a...change without a DNA double-strand break. But base editors aren’t perfect. They may be slow, can only target...this month. Another issue that arises for base editors is that the editing domain must induce the base...compatibility to Cas homologs. Evolving better base editors with the help of phages! The latest ABE from the...bacteria links the expression of gene III to base editor activity and editing speed and consists of the ...