...different Cas12a inhibitors. Some of these anti-Cas12a proteins have broad inhibitory effects on Cas12a...Caixia Gao’s lab has developed a new plant base editor from an evolved tRNA adenosine deaminase that is... is fused to a nicking Cas9. This base editor can be used for A•T to G•C edits in protoplasts or plants... Biology Find plasmids containing these base editors Spatially and temporally regulated AAV-based ...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics and a screening approach...

...zinc finger nucleases to transcription activator-like effector nucleases (TALENs). Our most popular kit... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these two...now have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene...be popular years after they were added to the repository, including beta-catenin reporter M50 Super 8x... GCaMP6 series of genetically encoded calcium indicators were the hot technology of the year in our collection...

...DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering... “Transcriptional activators are easy to expand or combine with different effectors. I’d encourage people...The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn...devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...expression levels reliably increased in vitro by a factor of 10 to 20. A flexible and modular optogenetics...

...NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused...after that, the lab published a paper on an RNA editor that allows C to U edits (RESCUE). We’ll walk through...translational machinery treats like guanine. This RNA editor would permit point mutations in RNA which could...nonfunctional. In their quest to build a robust RNA editor, the lab started with the Cas13 scaffold, testing...PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the...is too large to fit into adeno-associated viral vectors, they tested truncated ADARDDs to see if they could...edit multiple RNAs. It’s clear that CRISPR RNA editors will open new doors for scientific research - how...

...papers are analogous to mammalian viral vectors in that the vectors are replication-deficient. Thus, a very...like for organisms like C. difficile. In 2014, Citorik et al. and Bikard et al. developed sequence-specific...antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, they used 1-2 CRISPR...sequences found only in certain bacteria (Figure 1). Citorik et al. targeted beta-lactam and quinolone resistance... for bacterial CRISPR targeting. Phage-derived vectors carry CRISPR RNAs (crRNAs) targeting either genomic...suggest that targeting multiple resistance/virulence factors may further reduce this likelihood. The key problem...hardest-to-kill bacteria. In using phage-derived vectors to carry CRISPR, we’re turning the tables on the...

...Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our Blog...discovered the modular format of some transcriptional activators: that the DNA binding domain (DBD) and transcriptional...of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists took advantage... original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and the lacZ reporter gene...detectable reporter signals depend on a number of factors, including protein expression and correct folding... is to use a number of different bait and prey vectors. This has been shown to be as effective as using...transcription-activating function of a eukaryotic regulatory protein. Science. 1986; 231:699-704. PubMed PMID...

...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and...block out of focus fluorescence from reaching a detector or restricting fluorescence to specific excitation...light passes through a pinhole before reaching the detector. The pinhole is specifically placed to only allow...allow light from the focal plane to reach the detector and all light from above or below is excluded. ...the optics of a microscope before it reaches the detector. This means that two or more objects lying within...collected and projected back to a fast, sensitive detector such as a scientific CMOS camera. If plasma membrane...higher photon flux via the more efficient camera detector and lack of a light-reducing pinhole. By collecting...

....com. Mentors You may have an adviser, but do you have a mentor? There is a difference. A mentor is someone...that many graduate and post-doctoral programs lack any formal or mandatory professional development training... you may need to look a bit harder to find your mentor. This may be a connection you’ve made at a conference... Alternatively, it might even be a colleague. A mentor does not necessarily need to be someone a lot older...planning, has written a great blog on choosing good mentors for scientists. Continuing professional development...popular job site webpages. Importantly, find a good mentor that can share relevant experience and resources...

...Cytosine base editors with minimized off-target DNA editing The original cytosine base editor, BE3, induces...deamination. They generated these editors using the BE4max architecture. The new editors reported in this study ...the paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus ...ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and the ctrA sgRNA...lab developed new fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of ...editing by TET1 by co-delivering TET1 and protein factors for linking methylcytosine oxidation to DNA repair...

...place. As a curator of the thousands of amazing plasmid technologies contributed by our depositors, Addgene...plasmid" section Anytime you use a plasmid from the repository, we request that you cite it using both its Addgene...increases the visibility of both you and the original depositor. Our citations feature makes it so that, when ...reliability of our search results depends upon you, our depositors, giving us as much information as possible during...you to browse through the many plasmids in the repository, our scientists have curated a number of special...attention to the great variety of plasmids in the repository. And last, but certainly not least, we often ...Special Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...