...The most requested ubiquitin plasmids were pRK5-HA-Ubiquitin-WT and HA-Ubiquitin, and we now have 19 different...different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene had hit our...cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids you'll note that those are...Actually, these two plasmids from Didier Trono's lab are our two most popular plasmids of the past 10 years...influx of plasmids for reprogramming differentiated cells into pluripotent stem cells. Plasmids from Shinya...requested. Ubiquitin - One particular gene we saw increased interest in from 2008 to 2009 was ubiquitin. The...our stride. We shipped our 100,000th plasmid. A variety of plasmids continued to be popular years after...

...on Addgene.org CRISPR Plasmids – Protein Tagging CRISPaint Gene Tagging Plasmids 101 eBook Resources ...Resources on the Addgene blog Plasmids 101: Protein Tags Plasmids for Endogenous Gene Tagging in Human Cells ...cloned in-frame to the protein of interest for plasmid-based expression systems. Degron tags are one of...protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation. Adapted from Nabet...interacting ligands. After an E3 ligase facilitates ubiquitination of the Halo-tagged protein in question, it ...endogenously expressed SCF complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added in...TIR1 F-box associated E3 ligase complex will ubiquitinate the AID-tagged protein which will target it ...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        New toolkit...In plants, auxin binds to TIR1 of the SCF-TIR1 ubiquitin ligase complex, promoting its interaction with...have this response, they do contain the specific ubiquitin ligase pathway needed for AID. By tagging proteins...cells, which interacts with the cells’ endogenous ubiquitin ligase, the Kanemaki lab was able to translate... the Kanemaki lab has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins ...

...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...and studies of protein ubiquitination and degradation.   Find the CRBNmidi plasmid here!   Kroupova, A.... or want to swap out the constant domains, the plasmids for the heavy chain and light chain are also available...By Mike Lacy Several types of drugs target the ubiquitin E3 ligase cereblon (CRBN), including popular new...with the multiple myeloma drug mezigdomide and ubiquitination substrate Ikaros (IKZF1ZF2) (B). Image adapted...minimal background.   Find these C. elegans mScarlet plasmids here!   Cao W.X., Merritt D.M., Pe K., Cesar ...disruption in Mycobacterium abscessus using a dual-plasmid CRISPR/Cas9 system By Aryana Khosravani Deborah...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... our hot plasmid articles from April 2018. If you'd ever like to write about a recent plasmid deposit ...an E3 ubiquitin ligase, is the basis of TRIM-away. In normal physiology, TRIM21 recruits ubiquitin and ...sign up here. No time to read? Listen to our Hot Plasmids Podcast! TRIM-Away: Targeted endogenous protein...your protein knockdown experiments! The TRIM-Away plasmids can be found here. Clift et al. Cell. 2017. PubMed...Endosomal pH Sensors podcast segment The first Addgene plasmid deposit from Professor Stefan Hell, who won the...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...head over to our CRISPR Plasmids and Resources page.  Plasmids for delivering CRISPRi components by a single...inducible degron system for C. elegans New CRISPR plasmids New items from the viral service Epigenetic ... Haynes at Emory recently contributed a set of plasmids to the repository that will help epigenetic researchers...researchers custom design their own engineering plasmid tools in about a week. Taking advantage of Golden...

...1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...Resources on Addgene.org Browse all CRISPR plasmids CRISPR plasmids for protein tagging Find validated gRNAs...degradation tag (dTAG) system, to co-opt the endogenous ubiquitin proteasome machinery to degrade any protein. We...formation between the FKBP12F36V-fusion and the ubiquitin proteasome machinery through binding with cereblon...Detailed protocols describing the use of these plasmids are available here. The Methods section of Nabet...cloning procedures used to construct the deposited plasmids. Molecules: We synthesized dTAG molecules by conjugating...terminus should be tagged, the dTAG lentiviral plasmids can serve as a rapid means of comparing functionality...

...example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but so are lentiviral vectors...reason, the top requested plasmid of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’...pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin ligase, cellular senescence...Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances...’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000 times (over 800...to check out the Trono Lab website. If you have plasmids you’d like to deposit with Addgene, be sure to...Who knows, maybe yours will be the top requested plasmid of 2018! Additional Resources Quick Guide to ...

...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible.... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...expression mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  As the name implies, this...General expression mRNA Plant promoter from maize ubiquitin gene Constitutive  Gives high expression in plants...

... best plasmid for your research can be a daunting challenge. With over 80,000 deposited plasmids from ...through to the plasmid search results: More than 200 different GCaMP containing plasmids are available...search tool lets you access plasmids from journal articles. From paper to plasmid Authors regularly indicate... to include specific plasmid ID numbers. If you know the ID number for a plasmid, your straightforward...page to see the plasmids deposited by our own CSO. But what if you don’t have plasmid ID numbers, or you... Not every plasmid search will start from a publication. You may have an idea for a plasmid that would...exact plasmid you need. The ability to apply a wide range of filters isn’t new to Addgene’s plasmid search...

...localization, or protease protection. Thus far Plasmids 101 has covered GFP and its related fluorescent...crystallizing it or testing its function. Small ubiquitin-related modifier (SUMO) can help with folding ...stability activity, can be readily purified using plasmids found in this paper (available at Addgene).   ...interest: Which Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent...   plasmids 101...

...transforming both fusion plasmids, or mating two single haploid yeast fusion plasmid transformants into a ...reporter system (e.g. the Gateway-compatible bait Y2H plasmid pEZY202  from Yu-Zhu Zhang’s lab) is an example...requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References 1. Casadaban...Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our...

...labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile...compared to transgenic animals (17).  These viral plasmids are available through Addgene.  The combination...University of Miami Miller School of Medicine. Find plasmids from the Lee Lab here. Pantelis Tsoulfas is an...University of Miami Miller School of Medicine. Find plasmids from the Tsoulfas lab here.     References: H...

... Can I use my favorite plasmid with this protocol? Maybe. Not all plasmids will work for the one-pot...strands of the plasmid will be nicked during the first BbvCI digest and the whole plasmid will be degraded...those in the plasmid. Again, if they are in the opposite direction, both strands of the plasmid will be nicked...map of your plasmid. If the sites are in opposite directions, you just need to use a plasmid with BbvCI...Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Find Plasmids from the Whitehead Lab Use.... This technique relies on the generation of a plasmid library that expresses all desired variants of ... a selective pressure winnows the pool down to plasmids expressing variants with optimal function. High-throughput...

... 35S) or ubiquitin promoters (Belhaj et al., 2013). Addgene carries Cas9-containing plasmids for knockout... consists of a Ti plasmid, into which the GOI is inserted, and a Ti "helper" plasmid, which contains Vir...for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and the...plant-specific modifications to commonly used CRISPR plasmids are necessary to use the CRISPR system in plant...expression of both Cas9 and the gRNA off of the same plasmid. Delivering CRISPR components to plant cells Once...and insertion into the plant genome. B) The Ti plasmid and helper are transformed into agrobacterium tumefaciens...and exposed to plant cells. The region of the Ti plasmid between the T-DNA borders (yellow) are recognized...

... for panel design. Find streptavidin plasmids and biotin plasmids at Addgene!     Figure 1: Indirect...chemokines; enzymes like kinases, phosphatases, and ubiquitin ligases; and many more. This process, however,...Addgene Addgene's Antibody Guide Addgene's Antibody Plasmid Collection Resources on the Addgene blog Antibodies...

...a specific tissue. XFPs are introduced using a plasmid construct called Zebrabow-GateDest containing an...YFP. After crossing onto a tamoxifen-inducible ubiquitin background, each cell in the mouse that expresses...introduced in vivo either by electroporation of plasmids, or using AAV vectors. The Imai lab also developed...

...You’ve gotten the plasmid encoding your protein of interest from Addgene, transfected it into your target...from a variety of vendors and you may even find plasmids encoding antibodies from Addgene that you can ...phosphorylation, acetylation, methylation and ubiquitination. For example, to test if a protein is phosphorylated...

...Neurosci 2016. PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...stocks in a single plate with every plasmid or as individual plasmids shipped as agar stabs. Iverson, et...

...because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How to Design ...straightforward in easy-to-transfect cell lines where plasmid dosage can be tightly controlled. This is also ...Resources on Addgene.org Find fluorescent protein plasmids and resources Visit our Molecular Biology Reference...regulate numerous cellular functions, including ubiquitinoylation, kinase signaling, integrin signaling, and...