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TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m.... 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...
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Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....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... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog PostPublishedJuly 12, 2021, 1:15 p.m....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... -
Viral Vectors 101: Chemogenetics
TypeBlog PostPublishedJuly 11, 2023, 1:15 p.m....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... -
Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog PostPublishedNov. 7, 2017, 1:59 p.m....immunogenic profile. AAV was first discovered as a contaminant of adenovirus preparations. It is an icosahedral...untreatable until 2008. Following a single dose of subretinal administration of rAAV2 carrying the human gene... responses to adeno-associated virus and its recombinant vectors." Gene therapy 10.11 (2003): 964-976.... -
Tips for Technical Support Calls
TypeBlog PostPublishedSept. 20, 2016, 2:39 p.m....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 PostPublishedMarch 15, 2016, 2:30 p.m....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 PostPublishedJuly 23, 2015, 2:30 p.m....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 PostPublishedOct. 22, 2014, 1:57 p.m....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... -
Hot Plasmids: Summer 2025
TypeBlog PostPublishedAug. 28, 2025, 1:15 p.m.... 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... -
Transferable Skills Guide: Managing a Team
TypeBlog PostPublishedJan. 30, 2018, 2:42 p.m....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... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog PostPublishedJune 25, 2024, 1:15 p.m....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... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog PostPublishedFeb. 7, 2019, 2:16 p.m....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 PostPublishedApril 26, 2022, 1:15 p.m....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 PostPublishedAug. 10, 2021, 1:15 p.m....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... -
Twenty Years of Addgene Sharing: CRISPR
TypeBlog PostPublishedApril 18, 2024, 1:15 p.m....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 PostPublishedFeb. 23, 2016, 3:30 p.m....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... -
Tips for arabidopsis transformation
TypeBlog PostPublishedOct. 25, 2018, 1:23 p.m....are binary gateway vectors and can be used for promoter swapping and generating N and C terminally tagged...4. Nakagawa, T. et al. (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation ... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....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... -
Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!
TypeBlog PostPublishedDec. 3, 2019, 11:38 a.m....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....