...transferred between cells. At the time, these extranuclear agents of heredity were thought of as everything...first plasmid “cloning” experiment. Dr. Cohen and colleagues treated a tetracycline resistant plasmid, pSC101...topology, and cloning and gene expression. Just released: Addgene's Plasmids 101 (2nd Ed.) eBook And now...and more pictures and pro tips.  Happy reading! Learn more about the history of plasmids and cloning: ...

...transduction.) Compared to CCP transduction, iTOP is at least four times more efficient in primary fibroblasts... gRNA, which subsequently enter the nucleus and cleave the specified target. Figure adapted from D’Astolfo...expression is by nature transient, which may lower nuclease toxicity or the potential for off-target effects... Biol. 2000 Jul;10(7):290-5. PubMed. Is VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat...

...embryo—even if it’s not necessarily via CRISPR/Cas9. This leaves us with the second critique. Absent of a broad ...intended parents is “rehoused” in a separate, enucleated zygote with healthy mitochondria, with the intent...Nuffield Council’s report is that when it became clear that this assistive reproductive technique was feasible...featuring a clinical pregnancy, was rejected by at least one academic journal for exactly the critiques leveled...

...accessible to everyone but these projects could also lead to novel yeast creations that may not be pursued... the different samples. The goal is to not only learn something about the microbial diversity in different... to find a community lab in a city near you. To learn more about what we’re up to or how you can help ...Disease & Open Science: Interview with Darrell Kotton Learn How Plasmid Sharing Speeds Science Resources at...

... work to enable future discoveries. Read on to learn more about Xin and Fatima's work and keep checking... work at the 2017 Winter qBio Meeting. You can learn more about Xin’s work with photoswitchable components...enzyme breaks down the HMO of interest, the lactose released activates expression of GFP from the lac operon...Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent Proteins...

...viral vectors. Here are some of the AAV we have released in the last few months. You can also browse our...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency...CRISPR-Csm presents an attractive alternative for nuclear-RNA targeting, where siRNAs are ineffective and...expression changes of non-target genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted...antibody per target just isn’t enough and we recently released Anti-GFP [N86/8R] as another option for targeting...

...interesting at an event. Or, perhaps you have a colleague who seems fun and helpful who you’d like to get...connections. I mention this because I want it to be clear that it is completely impossible for me or anyone...contacts over the years in many different ways. This leads me to my last tip: It’s usually not worth connecting...networking Attending a conference soon? Get tips here Learn about informational interviews ...

...it unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent problem... perform an optimization-scale infection and at least one screening-scale experiment achieving the gRNA...the publications associated with these libraries. Please note, the pooled libraries we distribute in lentivirus...or are thinking about using our viral services. Please send any questions or comments to help@addgene....

...night of the iPSC symposium, a panel of thought leaders in the field--including Shinya Yamanaka, George... of ethics can be daunting because they have no clear cut answers, I applaud the stem cell community for...Disease; and Kevin Eggan presented work that is leading to promising clinical trials for ALS.   I have ...that!  Additional Resources on the Addgene Blog Learn about Efforts to Fight Rare disease with iPSCs Read...

...ever like to write about a recent plasmid deposit please sign up here.  Listen to the hot plasmids episode...Vellai lab has created an improved RNAi vector that leads to significantly stronger phenotypes than standard...incorporation of non-specific nucleotides into the dsRNA leading to increased RNAi efficiency. Find the T444T ... forms an active LacZ tetramer.  This tetramer cleaves chlorophenol red-b-D-galactopyranoside on the assay...

...Additional Resources on the Addgene Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our...of 2016. The top viruses of 2016 were (drumroll please)... Top Lentivirus: lentiCas9-Blast Most of Addgene... to perform unique editing tasks, classic Cas9 nuclease, which introduces double stranded DNA breaks, ...

...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn more about lambda... novel genetic variants in bacteria. Read on to learn about the key components of EvolvR and its potential...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   ...

...are some of the AAV and lentiviral preps we have released in the last few months: Lentiviral preps for ...coming soon! These vectors listed below should be released at Addgene in the next 6 months, pending quality... particular catalog item is available for order please email [email protected] with the vector name and...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...fluorescent reporter assays to quantify targeted Cas9 cleavage, and repair through homology directed repair (...while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using the supplied... seamlessly remove the integrated GFP reporter leaving you with just your desired genomic edit.  Find ...using GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting...

...collection in your experiments and published your data, please fill out this form and we’ll add your citation ...combinations of WT, codon optimized, untagged, cleavage sites (thrombin and TEV), and C-terminal tags ...downstream experiments and offer flexibility in tag, cleavage, codon usage, and expression system. Mammalian...Addgene blog Read all of our COVID-19 blog posts Learn about other COVID-19 plasmids available at Addgene...

...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this...of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-...anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin... editing, while gene editing can be done via a nuclease-active CasMINI. This small yet powerful system...

...option, in aliquots no smaller than 10uL. You can leave them at room temperature when you’re working with...illustrious lab career, you will likely accidentally leave your antibody out on the bench overnight. Do not...this post, the conjugates, or signaling molecules! Please keep in mind that this is just a brief introduction...data, you’ll want to pick fluorophores with the least spectral overlap between them whenever possible....

...the nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during cell...there – most of which we would all like to steer clear of! However, a subset of viruses are commonly used...which ones could be helpful to you. Interested in learning more about a specific virus? We have you covered...

...ever like to write about a recent plasmid deposit please sign up here. Here's what you'll find in this...a continuation of that work, they have found at least two variants that display increased neuronal expression...microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system reports an ... less risky.   Fig. 3: Development of a cleavage-free dCas9-based gene editor using microbial SSAPs...