...containing these mutations have been deposited at Addgene. Plasmids deposited from Alejandro Balazs’s lab includes...including a handful of plasmids that have been in the repository for many years, has now grown into a collection...lentivirus with the SARS-CoV-2 spike protein. They deposited the plasmids used for this study at Addgene which...can be completed in just 45 minutes. The team deposited the reverse transcriptase, protease, and Bst polymerase...  Drew Endy’s and Philippa Marrack’s labs also deposited plasmids for producing COVID-19 diagnostic enzymes...cathepsins play a role in SARS-CoV infection and deposited plasmids for the expression of cathepsin S, L,... of these proteins with human proteins. Their deposited plasmids expressed 26 of the 29 SARS-CoV-2 proteins...

... sequence or a sequence provided by the depositor. Depositing labs often provide insert sequences or annotated...by the depositing lab, we call these “quality control (QC) issues.” We then ask the depositing laboratory...take when performing quality control of newly deposited plasmids is to align the NGS result to a reference...laboratory to review the discrepancies. If the depositing laboratory confirms that these differences are expected...information about our sequencing results in the Depositor Comments section at the bottom of the plasmid .... In general, our QC scientists work with the depositing lab to develop a PCR strategy suitable for the...hope to make available as part of our OpenBio Repository sometime soon. Stay posted!     References ...

... versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and K27 (pcDNA3-K27...The K9 reporter uses the HP1 chromodomain to recognise lysine 9 methylation, while the K27 reporter uses... methylation in living cells. Essentially this reporter construct consists of one substrate (the histone-derived...-K27 histone methylation reporter) positions on histone H3, which are known to be involved in repression...methylation of these sites was controlled, the reporter consistently showed significantly different FRET...states. Thus, this system is a reliable tool in reporting methylation status of these sites. This new technology...Alice Y. Ting. "Genetically encoded fluorescent reporters of histone methylation in living cells." Journal...

... added a new feature for depositing labs to its website, allowing depositors to download their plasmid...information can be of personal interest to the depositor, but it can also be valuable data to share in ...grant proposals or to demonstrate impact in annual reports. While the raw data is, of course, interesting ...is launched, it will show my data by default. Depositors can download their CSV file from Addgene by following...Dashboard After the upload, the name of the depositor and the date of downloading the CSV is shown (...and the source code is available in the Github repository (you can find the link in the upper right hand...dashboard useful when analyzing and sharing your own depositor metrics!  About the Author Joachim Goedhart is...

...vector (deposited by David Root of the Broad Institute) has consistently been one of the repository’s most... to upload protocols when they deposit their plasmids to the repository. These protocols show up on the... and you decide to share them using Addgene’s repository, you can also share your associated protocols...“Supplemental Document”: gRNA_Cloning Vector (deposited by George Church’s lab). Addgene has also cataloged...the benefits and how you can get started on our deposit page. Protocols.io Website: http://protocols.io...the software aims to be a life science protocol repository. The software can be used with the ZappyLabs ...Protocol Exchange (NPE) is an open online protocol repository hosted by the Nature Publishing Group. Like the...

...combination with that reporter, you can infer its presence through detecting the reporter's fluorescence. Another...labeling proteins is to fuse them with a fluorescent reporter, such as GFP or mCherry, through genetic modification...combination of antibody-fluorophores, fluorescent reporters, and/or dye stains used in your flow cytometry...mentioned previously, you can also use fluorescent reporters, such as GFP, and fluorophore-conjugated antibodies...compare the excitation/emission spectra of your reporter with the ones from your fluorophore labels and...encodes for eGFP, which serves as a transduction reporter. Note that eGFP is not fused to CD45, but is separately...for having a separately expressed transduction reporter is that not every transduced cell will necessarily...

...on creating a repository that made it easy to share plasmids, both for people depositing plasmids and ... materials in our repository. We started to increase accessibility to our repository by making it easier...to — so far, we’ve shipped materials from our repository to an amazing 111 countries (Figure 1)! But while...distributor allowed us to share materials and accept deposits from scientists in China. They were also able ...strides in increasing the accessibility of our repository, from physical materials to useful information...changes. It was only after creating a plasmid repository and shipping internationally, after all, that... addgene.org. Currently, we’re evaluating our depositor process to identify and reduce barriers for scientists...

...information from requestors and depositors, and identifying trends in our own repository data, giving us significant...been finding and sharing stories unique to our repository and resources. As editor of Addgene’s eBooks ...order high quality viral vectors from the Addgene repository can save researchers significant time and removes...Cumulative AAV preps available in the Addgene repository each year from 2016–2023. B) AAV preps distributed...serotypes. In fact, the 879 viral vectors in our repository are produced from only 413 plasmids containing...different types of tools scientists were using/depositing. Our viral vectors data is less amenable to such...GCaMP-encoding viral vectors available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene...

...potential new deposits, and to meet with scientists throughout the world to encourage them to deposit. However...for in the repository? Let us know! You can notify us to reach out to potential depositors in two ways...reach out to will end up depositing, we hope this campaign will expand the repository in ways that will be...help us reach out to the corresponding potential depositors. 2. Tweet at us! Tweet @Addgene with #suggestaplasmid...

...use of both when making plasmids. Use repositories Repositories such as Addgene help share materials quickly...After the great Plasmid Debate of ’18, Blue Flame depositor John Doench calculated the time to make a plasmid...in a 96-well (or 384-well) plate! How much do repositories help?   Designing precise calculations for any...clear that community engagement with and use of repositories like Addgene provides large-scale savings. How...and the 2018 numbers for consumable costs, our depositors have helped us save the scientific community ...on Addgene.org Join the community of Addgene depositors here Check out our special collection pages to...find plasmids Search for a plasmid in Addgene’s repository ...

...a UAS-driven luciferase reporter construct. In the absence of GFP, no reporter output was observed in ... and a reporter construct into the mouse retina. GFP expression robustly activated the reporter gene TdTomato...Addgene, we love GFP, and we’re always excited when depositors find new ways to make this workhorse protein ...and Cre-DOG, respectively, repurpose popular GFP reporter lines for more sophisticated experimental manipulations...GFP. They then tested T-DDOGs with two mouse GFP reporter lines; again, TdTomato was seen only in cells ...tested in retinal electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, ...

...fluorescent reporter, directly couple silencing of your gene target to silencing of the reporter. This is...fusing your gene target sequence to a fluorescent reporter sequence (discussed below). Examples of these ...contains a target sequence against the fluorescent reporter in an inverted repeat orientation. Therefore, ...sequence against your gene(s) of interest and the reporter. This approach greatly enhances experimental throughput... observable phenotypes quantified. Fluorescent reporter-based RNAi constructs have been designed and widely...processes to isolate RNAi plants. With a visual reporter of RNAi, phenotypes, such as morphological parameters...identified as undergoing RNAi through the fluorescent reporter and manually extracted. This process is tedious...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...articles provide brief summaries of recent plasmid deposits and we hope they'll make it easier for you to ...you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in...in a protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed ...introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim’s lab engineered...library that targets human epigenetic genes was deposited by Kivanc Birsoy. Find the library here A new...  Find the plasmid here Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa system...

...unique skills and strengths (you will likely have reports with different levels of experience), find out ...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...itself is counter-productive. I always assume that reports want to succeed at work and providing feedback ...feedback is needed for that. In many cases reports are not aware of issues until you bring them up. I wouldn’...wouldn’t go as far as saying that your reports will be grateful for the feedback, but at least you can start...someone’s hard work and reward a job well done. Your report will (needs to) feel valued. Talk to your manager...

...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ... using IRES elements. Further, scientists have reported lower expression of the downstream cistron due...eukaryotic cells, and, although some scientists report close to 100% cleavage with some of these peptides...that already has the multicistronic element and reporter cloned in. In these plasmids you would simply ... 2A element (depending on the placement of the reporter gene). Addgene's collection offers a variety of...multicistronic element(s), or you could replace one of the reporter genes above with your gene or genes of interest...equivalent levels upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's...

... our viral vectors repository this year (and it's only March!)  In our repository, you'll find the latest...viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes AAV-CAG-jGCaMP8s-...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the 21...vectors that can constitutively express JEDI-2P, deposited by the St-Pierre lab. Also of interest is the ...check out Addgene's Data Hub to view viral vector reports from other users. And don't forgot to use the link...

... people usually report? Addgene and other viral vector manufacturing facilities report the physical titer...cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-infectious ... However, for recombinant AAV2, the same study reported a physical-to-infectious particle ratio of 50:...(“full”) (Zeltner et al., 2010), while another reported that only 20% of recombinant AAV2 particles were...vector preparation protocol, and we are pleased to report that over 95% of viral capsids in representative...AAV aliquots. Our titer values (circled in red) report physical titer, which is measured by qPCR. ...nonsense to publication quality data." Scientific Reports 7 (2017). PubMed PMID: 28546538. PubMed Central...

...molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can tell us about cellular events...higher apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general...protein-protein interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since...fluorescence of co-transfected EGFP. e Originally reported as a monomer16, IFP2.0 was later found to be a...E., Sivanesan, D. & Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein...Filonov, G.S. & Verkhusha, V.V. A near-infrared BiFC reporter for in vivo imaging of protein-protein interactions...evolution approach applied to fluorescent voltage reporters. Nat Chem Biol 14, 352-360 (2018). Yu, D. et al...

...research. If you’d ever like to deposit a kit, please contact us at depost@addgene.org. ...of the many useful kits you can order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System...library by in vivo functional screen." Scientific reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted...strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013. These terminators... an Orthogonal Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1 (2015): 31-37. pCri...quantitatively determine allele frequencies across timepoints or populations. This kit consists of a 48 plasmid...fusions. Some applications of PUF protein fusions reported in the literature include RNA detection, RNA cleavage...

...data repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for ...commercially available sources, as well as those deposited with Addgene. NCBI BLAST: Compares nucleotide ... Calculator app for iPhone Reagent and Data Repositories These links can help you find experimental and...for Cas enzymes.  Open repository of CRISPR screens Cellosaurus: A knowledge resource for cell lines IBEX...IBEX Knowledge Base: A data repository for validation data on IBEX reagents.  mAb3D atlas validated antibodies...human, and marmoset brain IHC.  FPbase: A data repository for fluorescent proteins and their properties...