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TypeBlog Post...dCas9. Within a few rounds of cell division, the original methylated strand is diluted to undetectable amounts...dCas9 so that it may later be excised by Cre recombinase. Addgene has several great options: pLV hUbC-...L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function...
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A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog Post...Originally published Apr 30, 2020 and last updated Jul 12, 2021. What started out as a small collection...used for this study at Addgene which include a C-terminal deletion in the spike protein to improve expression...pseudoviruses to determine how effective sera from vaccinated individuals are at neutralizing the different... -
Design Tips for Prime Editing
TypeBlog Post... Editing, and that meant rereading many of the original papers reporting various prime editing tools. ...your curiosity, we encourage you to check out the original papers for more details. You can find many software...prime editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652... -
Viral Vectors 101: Chemogenetics
TypeBlog Post...AAV serotypes, and the ever-growing variety of recombinase systems. Picking Ligands Picking the right ligand...ligand co-developed by Bryan Roth's and Takafumi Minamimoto's lab called deschloroclozapine (DCZ) shows promise...our chemogenetics poster! This blog post was originally written by Maya Peters Kostman in May 2020 and... -
Tips for Technical Support Calls
TypeBlog Post...majority of our plasmids and pooled libraries were originally described in a peer-reviewed scientific publication... our thousands of depositors. That means their original articles are your best bet to figure out how to...asking to avoid unrealistic expectations. And finally, provide feedback. If you are unsatisfied with ... -
Tips for Titering Your Lentiviral Preps
TypeBlog Post...PCR product that comes from amplification of the original plasmid which can over-inflate results. A packing...vectors carrying fluorescent markers are used as it eliminates the need for antibody staining. Check Out Our...into a target cell’s genome. This approach can discriminate multiple integration events, reducing the possibility... -
Experimenting in an Open Source lab: from CRISPR to Cats
TypeBlog Post...researchers to use. Although we haven’t yet made our final target decisions, we intend to focus on motor neuron... a new project at a makerspace is finding the financial resources to pursue it. Since makerspaces are ...equipment for new uses. We’ll probably never use the Illumina GAIIx we recently acquired for sequencing again... -
How-to: 5 Steps to a Great Panel Discussion
TypeBlog Post...This post was originally published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates...members all say the same thing and one panel member dominates the discussion? Or maybe the one that goes way...front of a crowd. The moderator will then develop a final list of questions for the actual event, based on... -
Transferable Skills Guide: Managing a Team
TypeBlog Post...don’t get done if there is not a clear deadline. Finally check-in regularly, make yourself available, and...that one on one discussion can be confidential. Finally, provide timely feedback. We often associate feedback...manner. If someone’s performance or behavior is inadequate, you need to address it as soon as possible for... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...generated info for your own purposes, or delete the originals if desired. It works as a whole or in part. For...ranging between 0 and 1. The cutting frequency determination (CFD score) is used to measure specificity of...re-binding. Lastly, the sequence inserted by HDR may inadvertently re-create a match for the active guide. With... -
PCR Applications for Copy Number Variation Assays
TypeBlog Post...increased sensitivity allows for a more confident determination of copy number, albeit within a range of RQ ...trends? How confident do you need to be in your determination to move forward with your experiment? Complexity...real-time polymerase chain reaction methods for the determination of amplification of Plasmodium falciparum multidrug... -
Polymerase Chain Reaction Overview and Applications
TypeBlog Post...the end product of PCR temperature cycling. The final PCR product is often visualized on a diagnostic ... dNTPs and other reagents to run low during the final cycles which would slow or stop PCR amplification...statistics to determine the concentration of the original template DNA in the sample. Addgene currently... -
Hot Plasmids: Summer 2025
TypeBlog Post... an approach called Simultaneous Tracking of Recombinase Activation and Clonal Kinetics (STRACK) to study...epithelial cells, liver, CNS (limited penetration), retina, hematopoietic cells, and immune cells. B) AAV11...postsynaptic AMPA receptor and is endocytosed, the Cre recombinase payload activates a floxed reporter gene (genetically... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog Post...Figure 1). Through this method, we found key determinants of prime editing efficiency such as the length....0 while accounting for some of the inherent inaccuracies of computational prediction models. In addition...efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org... -
Twenty Years of Addgene Sharing: CRISPR
TypeBlog Post...2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain the most popular deposited...Read our recent publication, “The Expanding Dissemination and Distribution Patterns of Diverse CRISPR ..., R., & Barrangou, R. (2023). The Expanding Dissemination and Distribution Patterns of Diverse CRISPR ... -
PITChing MMEJ as an Alternative Route for Gene Editing
TypeBlog Post...active during M and early S phases, when HDR is inactive, and the balance of NHEJ, HDR, and MMEJ repair...longer the homology, the more efficient the recombination, but with longer homology arms comes more time...which has been difficult to modify due to NHEJ’s dominance over HDR in this species. As CRISPR technology... -
Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!
TypeBlog Post...labs and institutions. For example, search for “Melina Fan” and you’ll be able to go directly to a page...In this example we’re going to be looking for recombinant monoclonal antibody plasmids from the James Trimmer... ‘Vector Types’ I selected ‘Lentiviral’ which eliminates most of the plasmids designed for other uses.... -
SciComm: Taking Science to Elementary and Middle Schools
TypeBlog Post...scientist. When our elementary school needed a new coordinator for the annual Math & Science Night, I took the...It’s through the PTA that I recently started coordinating the annual Math & Science Night, a chance for...contacting our school district’s science curriculum coordinator with minimal luck, but eventually the 8th grade... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...fragment to the N- and C-terminal end of each PIP and empirically test which combination provides the best fluorescent...cellular functions, including ubiquitinoylation, kinase signaling, integrin signaling, and transcription... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog Post...T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under conditions commonly...Methods Matter: Standard Production Platforms for Recombinant AAV Produce Chemically and Functionally Distinct...Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent...