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...science stories. Here are some of our favorite spooky science stories. Six spooky science stories Zombie...Plasmid pumpkins, team costumes, and spooky science stories. Yes, please! At Addgene, we love finding ways ...Vampire bacteria Vampirovibrio chlorellavorus is a predatory bacteria that attaches to the cell wall of green...forcing it out of hiding to become exposed to predators. Hungry birds spots the “caterpillar” and then... "Back from the dead; the curious tale of the predatory cyanobacterium Vampirovibrio chlorellavorus." ...

...ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have...Testing AAV inventory for the entire collection! We are expanding our entire control AAV inventory to give...Calcium Sensors). We are expanding the inventory of these vectors to more serotypes. We currently package...GCaMP7s. See all our biosensors inventory, and check out our latest vectors below: pGP-AAV-syn-jGCaMP7f-...addition to our inventory of Cre-dependent GABA sensors. See all our GABA sensor AAV inventory here. pAAV.hSynap.iGABASnFR...support by growing our inventory of recombinases and recombinase-dependent vectors. We recently released...pENN.AAV.hSyn.Cre.WPRE.hGH (105553-AAV5) Dre vector AAV phSyn1(S)-DreO-bGHpA (50363-AAV5) Flp vectors pAAV-EF1a-mCherry-IRES-Flpo...

...guarantee certain rights and protections for depositor and requestor alike. That’s why Addgene works closely...closely with TTOs to help protect our depositors’ intellectual property, and to help requesting institutions...California Institute of Technology Cold Spring Harbor Laboratory  Dana-Farber Cancer Institute  Fred Hutchinson...

...ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have...pEF1a-DIO-FLPo-WPRE-hGHp Other new viral vectors New optogenetics inhibitors Plasmid Serotype Name 99039 ...AAV1 pAAV-ihSyn1-DIO-tTA Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...few months. You can also browse our entire AAV inventory. Our new AAVs include: Control AAVs for serotype...always excited to add new control AAVs to our inventory since we know that scientists need these tools...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid...

...ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of the AAV we have... used in conjunction with optogenetic activators and inhibitors that respond to blue light. We recently...GABAergic interneurons. These vectors can also be used as the control vectors for experiments encoding a ...pAAV-hSyn-mCherry at Addgene. Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...far in 2019. You can also browse our entire AAV inventory! Our new AAVs include: pGP-AAV-syn-FLEX-jGCaMP7b-WPRE... Addgene. GABA sensor AAV GABA is the primary inhibitory neurotransmitter in vertebrates, but methods ...

...block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial constitutive...research purposes, retroviral vectors have been developed into ‘split-vector systems’ by separating viral...and expression of these vectors when used in experiments. Retroviral vectors make up a large toolbox ...Gamma-retroviral vs. lentiviral vector systems Gamma-retroviral vectors have a relatively high transduction... many more useful viral vectors available at Addgene: Follow our viral vector blog posts for updates on...tips for using retroviral vectors in your experiments. Gamma-retroviral vectors Popular gamma-Retroviral...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...

... 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...

... adapted them for use in the laboratory. The discovery of GFP The story of how fluorescent proteins like...Aequorea victoria (Figure 1) in molecular detail. On the underside of the “umbrella” of A. victoria there...turns out, he was right! He began to study the A. victoria gene responsible for encoding GFP, and, in 1992...sequence. Soon after that, in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms...studying it grew. In particular, studies in the laboratory of UCSD biochemist Roger Tsien resulted in much...

...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...

...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 ...

...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...

...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...

...expression vectors for insertion of custom gRNA target sequences.... CRISPR Empty gRNA Vectors CRISPR Plasmids...Plasmids: Empty gRNA Expression Vectors gRNA and Cas gRNA Only Multiplex Resources CRISPR requires you .... Addgene has a large collection of empty gRNA vectors. Explore our most popular plasmids for expression...tables below. Looking for non-targeting control vectors? Search our catalog for control gRNAs . All-in-...All-in-one Plasmids (gRNA and Cas) ID Plasmid Vector Type Promoter Selectable Marker PI Publication Return ...Return to top gRNA-only Plasmids ID Plasmid Vector Type Promoter Selectable Marker PI Publication Return to...

... CRISPR CRISPR History CRISPR History and Development for Genome Engineering ...Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I (Cas3) Type II ...technologies that do not rely on NHEJ or HDR: Base editors : Base editors directly convert bases without creating...The first base editors converted cytidine to thymidine; newly engineering base editors convert adenosine...various CRISPR-associated (Cas) genes to not only store a record of invading phages but also to destroy ...two large classes based on the structure of the effector complex that cleaves the genomic DNA. Type II ...adenosine to guanosine. RNA editors : Cas13 fused to the ADAR2 deaminase convert adenosine to inosine on RNA...

...Viral Vector Packaging Service Penn Vector Core Transfer Viral Vector Packaging Service: Penn Vector Core...AAV vectors to the academic research community. The Penn Vector Core produced viral-based vectors for ...Addgene's viral vectors See Addgene's Current AAV inventory , which includes all viral vectors transferred...Transfer The University of Pennsylvania Vector Core (Penn Vector Core) and Addgene previously partnered...a viral vector with the Penn catalog ID: Based on low demand, production of some viral vectors have been...See the list of discontinued viral vectors . The following vectors are now available for distribution ...Optogenetics Baljit Khakh Discontinued Penn Vector Core Viral Vectors (Plasmids still available, other serotypes...

...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...

...The hM4Di receptor is an engineered Gi protein-coupled receptor, one of the Designer Receptor Exclusively...DREADD receptors need to be located on the plasma membrane to be activated by the DREADD actuator after...brain cells. Since the opsins or chemogenetic receptors used in these techniques cannot be directly visualized...of interest. In our studies, we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or...identify the expression of the opsin or chemogenetic receptor. However, in some cases the fluorescent signal...fluorescent protein with the opsin or chemogenetic receptor, through an IRES or 2A sequence. In this case,...subcellular compartment as the opsin or chemogenetic receptor. This may not be an important limitation in many...

...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...

...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 ...

...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...

... contributed  by Jesse S. Boehm, the Associate Director of the Cancer Program at the Broad Institute of... plasmid toolkit, the Broad Institute Target Accelerator Plasmid Collection, that we hope researchers ... For many years, we’ve hoped that large scale laboratory-based functional experiments that classify which...and cancer phenotypes. Broad Institute Target Accelerator Plasmid Collections To overcome this first challenge..., we’ve developed the Broad Institute Target Accelerator Plasmid Collections, now widely available to ...Collection). In addition, we worked with many collaborators to aggregate a sequence-confirmed series of ...one assay per gene approach that has been the historical standard. Specifically, in our recent publications...

...passion, and dedication to sharing epitomized by the creators of these plasmids with the first annual Blue Flame...world choose to share their plasmids through our repository since we know biological resource centers, including...+ times Since we so sincerely appreciate our depositor’s dedication to scientific sharing, we wanted to...requested plasmids. Currently, there are roughbly 400 depositors worldwide who have one or more blue flame plasmids...will serve as badges of honor for our powerhouse depositors who have generously shared their reagents freely...community. Sharing is caring We’re thrilled that our depositors reap the benefits of sharing plasmids with us...us. One way we can gauge if people are happy depositors is if they deposit plasmids more than once. People...

...Co-founder/director of the non-profit foundation IMFAHE and its International Mentoring Program Director of Strategic...experience co-directing Mentor Circle Programs at BWH and providing career mentoring to several students ...successes. Although all mentorship relationships are valuable and highly recommended, a mentor with international...International Mentorship Program work? IMP Pilot Program 2015-2015 Scholars need strong proactive mentors who have...to their direct training role as mentors to Spanish mentees, mentors also facilitated the development ...Impact of the International Mentorship Program and the International Mentorship Conference In addition to...international mentoring conference held at the University of Valencia (Spain). At this conference, mentors and ...

...the sample for entry into Addgene’s repository. For long-term storage in our facilities, every sample is...used for long term storage (one is kept at Addgene and another is sent to a storage facility). The third...is the Laboratory Operations Manager at Addgene.  In addition to helping with daily laboratory operations...courier and then the journey of transformation and storage begins. Some samples are submitted as bacterial...through barcoded tubes that are scanned into our Laboratory Information Management System (LIMS), a database...sample can be tracked easily at each step in the storage process. After the original sample is barcoded,...different types of samples at Addgene - DNA sent from depositors, bacterial glycerol stocks, prepped DNA ready...

...Consortium: Nuclease Expression Vectors This page lists plasmid vectors designed to express nucleases in...transferred in-frame to any of these vectors using unique cloning sites. Vectors encoding wild-type FokI nuclease...This page lists plasmid vectors designed to express nucleases in different cell types. Zinc finger arrays...in-frame to these vectors. Plasmid... Plasmid Collections Zinc Finger Consortium Nuclease Expression Vectors Zinc Finger... zebrafish) pDW1775 (for expression in plants) Vectors encoding heterodimeric FokI nuclease domains: pMLM290...

...some good news with you: with contributions from depositors around the globe, we’re now sharing over 100,...100,000 different plasmids available from the repository (currently at 98,276 plasmids as of April 8, ...

...relies on the restoration of a frame-shift in mCherry that restores fluorescence to monitor APOBEC-Cas-9...with the BFP variant, a BE4am-Cas9 base editor, and sgRNA vectors for the BFP-to-GFP conversion and to target...Base editors create specific point mutations in the genome, but they’re inefficient compared to CRISPR...editing reporters that can help you do just that. Monitoring APOBEC and Cas9-mediated editing in real time...time The Harris lab created ACE, a reporter that monitors APOBEC (a family of cytidine deaminase proteins...that disrupts fluorescence due to a frameshift. Restoration of fluorescence can only occur through editing...results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using...

...including terminators and promoters, CRISPRi parts, selectable markers, transformation acceptor vectors, and...therapeutic potential for the split base editors. Find the split base editors at Addgene. Levy et al., Nature ...voltage indicator Article contributed by Shreya Vedantam Genetically encoded voltage indicators (GEVIs)...broad use. This kit is designed for use with acceptor vectors from the MoClo Toolkit from Sylvestre Marillonnet...of Phytochrome B interacting factor 6 (PIF6APB), respectively. The cofactor analog, phycocyanobilin (PCB...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...sign up here.        AAV delivery of split base editors Article contributed by Jennifer Tsang AAVs are ...

...you out. What is the Open Repository of CRISPR Screens? The Open Repository of CRISPR Screens (ORCS) database...This post was contributed by the Open Repository of CRISPR Screens. Imagine you’ve just discovered that...research. Fortunately, there’s a growing open repository of CRISPR screen data that might be able to help... resource developed by the Biological General Repository for Interaction Datasets (BioGRID) in order to...biologists. Figure 1: The BioGRID Open Repository of CRISPR Screens (ORCS) homepage. Screenshot...Screenshot updated 2024.   Using the Open Repository of CRISPR Screens BioGRID ORCS currently houses over ...metadata for each screen is captured by BioGRID curators, including the cell line/tissue type, enzyme system...

...or lentiviral vectors in our previous blog posts.  Viral vector injection Viral vectors allow a tight ...neuronal function directly such as receptor subunits or neurotrophic factors. Another option uses optogentics...is cloned into a viral shRNA expression vector and the vector particles are delivered into the brain region.... On the other hand, lentiviral vectors are superior packaging vectors for larger genes - they can hold...around 1012-1013 vg/ml (vector genomes per milliliter) and lentiviral vectors around 108 IU/ml (infectious...using viral vectors By modulating neuronal gene expression and activity, you can use viral vectors to study...viral vectors blog posts Find tips for preparing AAVs View our Beginner's Guide to Viral Vectors Resources...