We narrowed to 1,082 results for: TOR

... for depositing labs to its website, allowing depositors to download their plasmid request data, as a ...information can be of personal interest to the depositor, but it can also be valuable data to share in ...total requests, total countries represented by requestors, year of first reagent sent, year with most reagents...is launched, it will show my data by default. Depositors can download their CSV file from Addgene by following...Dashboard After the upload, the name of the depositor and the date of downloading the CSV is shown (...and the source code is available in the Github repository (you can find the link in the upper right hand...dashboard useful when analyzing and sharing your own depositor metrics!  About the Author Joachim Goedhart is...

...including the internet!) can vary depending on factors like socioeconomic status and geographic location...communication should be fun and engaging for the communicator as well as the audience, so pick a mode of communication...restrictions (or both!) will be a major limiting factor at multiple points. Before you embark on a project... convey enough information to tell a compelling story in an image or do you need a full exhibit? In a ....  Static. Can be difficult to convey an entire story or context via still images. Some things cannot ...information. Primarily useful for data-focused stories. In-person lectures, presentations, etc… Low... Low barrier to entry for both audience and communicators. Well-developed content creation platforms.  ...

...What do a viral vector production facility, food allergy testing lab, and the grad student down the hall... curves in their day-to-day work. Indeed, viral vector production facilities frequently use qPCR with ...with each step of the series diluted by the same factor. For example, Figure 1 depicts a 2-fold dilution... standard curves vary depending on a number of factors, including the user, equipment, assay incubation...measure.  Dynamic range depends on a number of factors, including the dilution series of the standard ...standard curve dilution series with lower dilution factors. If the unknown sample’s measurement is too high...assays you will be using ahead of time to see what factors may impact the dynamic range of the assay. Try ...

... the notation works? The + operator denotes intersection and the – operator denotes set difference. That...inclusive markers of regulatory regions in the genome, including enhancers, promoters, insulators, and more. I...proteins”) for NANOG, an essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds...of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus in the same family as...enhancers, lncRNAs, repetitive elements, repressors, insulators, and more. We know that this matter exists, but...criteria: no TTTTT allowed (this is a polymerase terminator), and no off-target effects for the identified...

... uses synthetic RNA regulators to control replication. Additionally, we refactored the natural system ...architecture that lets researchers plug in synthetic regulatory RNAs to create plasmids with new compatibility...pMB1, we used a previously-developed orthogonal regulatory RNA library to build six compatible SynORI plasmids... ORIs can be created this way with orthogonal regulatory RNAs designed computationally. Copy number control...ligand-responsive genetic elements, namely transcription factors and riboswitches, can be used to dynamically control...remains unclear, but the result suggests that refactoring the origin may have compromised a stability feature...

...keeping track of speakers. During your graduate or postdoctoral work there are loads of opportunities to create... concrete happen. You may have more willing collaborators than you realize. Being at a university provides...meetings and take minutes. Get to know your collaborators from other institutions personally on conference...relationship with the perspective of my potential collaborators in mind. Being at a university allows you to...with people at your organization. You can meet collaborators  for lunch, make a point of talking to them ... different countries). Here social media is a connector and virtual meeting place: You can follow interesting...as messages and keep you up-to-date on your collaborators’ professional developments without directly ...

...us. Use a well working vector system This allows you to quickly assemble vectors and test different ideas...MoClo system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo...to quickly screen and investigate transcription factor candidates and the regulation of genes in the biosynthetic... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...and a vacuum is applied with a vacuum bell or desiccator. Successful infiltration can often be seen by...system you can easily and quickly create customized vectors. For more information on MoClo for plants, check...

...content_id":null,"href":"https://www.addgene.org/depositor-collections/allen-institute-aav-enhancers/","href_with_scheme..."href_with_scheme":"https://www.addgene.org/depositor-collections/allen-institute-aav-enhancers/","type...interacting with proteins such as transcription factors and chromatin modifiers. These proteins recognize...within the enhancer sequence, then interact with mediator complexes and the RNA polymerase at the promoter...cell. Differential expression of transcription factors, which recognize and bind to different DNA motifs... of a fluorescent gene and packaged into viral vectors that can be administered systemically (retro-orbitally...reviewing this post! Dr. Ben-Simon is an assistant investigator at the Allen Institute in the department of ...

...-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School... Darrell recently joined the Addgene Board of Directors where he hopes to help keep us focused on our ...Darrell's goals as a member of the Addgene board of directors - increasing open source sharing and helping us...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...

...they do so is through G protein-coupled receptors (GPCRs), receptors that play many roles in signaling processes...which a donor fluorophore is used to excite an acceptor fluorophore. The difference for BRET is that the... between a luminescent donor and a fluorescent acceptor. TRUPATH uses an updated version of BRET called...constructing 14 luciferase donor Rluc8-Gα chimeras, four acceptor GFP2-Gγ chimeras, and two untagged Gβ constructs...non-uniform across the transducerome for a specific receptor-ligand pair. They further used TRUPATH to analyze...Mitaku S (2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters...

... referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may ...plasmid with its elements described below.   Vector Element Description Origin of Replication (ORI...the target gene. Vital component for expression vectors: determines which cell types the gene is expressed...Regardless, once the cloning steps are complete, the vector containing the newly inserted gene is transformed...Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of a chemical in a particular environment...about molecular cloning, how to choose a plasmid vector, molecular biology tools and references, and how...

... stands for Multiplexed Overexpression of Regulatory Factors. The Feng Zhang lab developed this library...libraries from the Samantha Morris lab use a combinatorial barcoding approach to observe clonal dynamics...nuclease-dead Cas9 is fused to a transcriptional activator or repressor to create a reversible effect. These...library of all human transcription factors, including splice isoforms, to allow comprehensive screening, cellular...Element. This library consists of broad host range regulatory sequences ready to be assembled with other elements...Additional resources on the Addgene blog Viral Vectors 101: Preparing Pooled Libraries Pooled Library ...

...upon an indicator they developed and published in 2010 for visualizing the transcription factor cAMP-responsive...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since the...Griesbeck is motivated by a particular interest in monitoring the activity and biochemistry of living neurons...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...while in principle adapting a FRET sensor for monitoring calcium versus zinc should require the simple... variants of mOrange2 and mCherry as donor and acceptor domains. He says, these new colors might now enable...

...Scientific Illustrator Plant Breeder Zoological Worker Conservationist Documentary Director Parasitology...skills include: Science Reporter Scientific Illustrator Research and Development Scientific Advisor...Lawyer Technical Writer Meteorologist Journal Editor Textile Technologist/Designer     Fieldwork...Bioinformatician Science Writer Grant Writer Journal Editor Science Marketer Software Developer Learn Science..., and how many jobs are normally available (as sectors such as forensics currently have too many applicants...hobbies and volunteer experiences, as well as work history can really show that you have some of the abilities...

..., the researchers did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform... study has just been uploaded to the preprint repository BioRxiv. This example clearly illustrates the...  Figure 2: When a researcher needs a collaborator for a specific task, Crowdfight will: 1) Find...researchers, such as postdocs and principal investigators. They also come from many different scientific...website. Once curated by our in-house team of collaborators, the requests were forwarded to our volunteer...IgnacioAmigoH     María Hernández-Sánchez is a postdoctoral researcher at Instituto de Investigación Biomédica...Biomédica de Salamanca (IBSAL) and one of the coordinators at Crowdfight. She is currently based in Spain...

...This will be highly dependent on vector dose. The color diversity in Vector Assisted Spectral Tracing (VAST...catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If so, you may...drop in transduction coverage compared to just one vector.   Q2: Do you think it is possible to make an efficient...this scenario, you may be best served by a dual vector, split intein approach.   Q3: What’s the best method...concentrated to ~1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011...Plasmids Adenovirus Guide Page Subscribe to Viral Vectors Blog Posts ...

...as several days. In 2015, the laboratory of long-time Addgene depositor Alice Ting used yeast display-...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...Recently the Vellai lab has created an improved RNAi vector that leads to significantly stronger phenotypes...for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted...Vellai lab’s new plasmid, T444T, contains T7 terminators adjacent to the T7 promoters to prevent the incorporation...Recently, the Sidhu group at the University of Toronto expanded the specificity landscape of the SH3 proteins...

...Oleispira antarctica, and the P. haloplanktis Trigger Factor), that would allow the stable folding and expression...compared to the E. coli strain expressing the empty vector. And thus was born ELSA - our happy lil’ cold-tolerant...of stakeholders from industrial and commercial operators, government agencies, active researchers, as well...interact with personnel from the industry and brainstorm with each other. The event also featured a seminar...The talk highlighted the importance of reagent repositories and the myriad facilities offered by them which...understanding of the societal ramifications of laboratory science, and how it may best be leveraged for...re always delighted to engage with potential collaborators and interested audiences. Please visit our team...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...regulate botulinum neurotoxin were also created. The depositors demonstrated the utility of light-mediated suppression...slices from mouse, and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids...acoustic reporter gene (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These.... 2018. PubMed PMID: 29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated ...microRNAs (miRNAs) are recognized as important regulators of post-transcriptional gene expression across...They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert site and a reporter...

...her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that they could...which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is an engineered protein complex for...information from the structure to guide and improve activator design,” she said. “Before it was a little like...Konermann recognized as an ideal spot to fuse the activators instead of at the protein’s C terminus. With ...activation complex consisting of multiple distinct effector domains that work together synergistically, modeled...melanoma cells resistant to treatment with a BRAF inhibitor. (The test was the perfect complement for one ...repressing or inducing individual transcripts. The activator (CRISPRa) sgRNA library uses the sunCas9 system...

...contributed by guest blogger Behnam Nabet, a postdoctoral fellow at Dana-Farber Cancer Institute. Targeted...provide distinct advantages over pharmacological inhibitors including rapidly depleting a protein-of-interest... target selectivity, overcoming resistance to inhibitors, and inducing prolonged biological effects. ...et al., Nature Chemical Biology for examples). Monitor neo-substrates Small molecule degraders may recruit...and IZKF3 are cell-type specific transcription factors with important roles in Multiple Myeloma), it is...studies (Nabet et al.).   Behnam Nabet, PhD is a postdoctoral fellow at the Dana-Farber Cancer Institute. ...dTAG-47 Weintraub et al. YY1 Is a Structural Regulator of Enhancer-Promoter Loops. Cell. December 14 ...

...via the secretory pathway, and then cleaved to generate an active form of the protein. To monitor autophagy... Mark Prescott. This system was developed for monitoring and analyzing autophagy of cytosol and organelles... samples treated with or without an autophagy inhibitor. While this method does provide a quantitative...the mammalian Atg8p homolog, can also be used to monitor autophagy, but under some conditions it aggregates...also possible to make Rosella fusions and thereby monitor autophagy of your protein of interest. Are you ...Cuervo, A. M., & Seglen, P. O. (2007). Methods for Monitoring Autophagy from Yeast to Human. Autophagy,3(3)...

...viral vector samples were distributed in 2018 and we are poised for more offerings. The viral vector team...don’t have worry or wonder if providing a viral vector service was a good idea. The scientific community...don’t have to worry about getting support from depositors. We regularly hear how much they see the value... 3.5 Addgenies strong. The Addgene Board of Directors is mature and poised to support our growth. Thanks...Addgene is about half of what is standard in most sectors. The Addgenies support a high collaboration, high...blog Subscribe to the Addgene blog Read our viral vector blog posts Learn more about Addgene Resources ...

... collisions you may encounter: vector, sequence, insert, trait, factor, expression, etc.There are also... Laboratory management software is not a requisite for a functioning lab, but it is for a scaleable lab...intentionally trivial, but they're here to help tell this story. In this first example to the right (# this will...about what steps are most likely to fail. Our laboratory manager would definitely be interested to see...this case, the physical considerations of the laboratory make object composition a smarter choice than...terms (plasmid, 5-prime, CRISPR, growth strains, vector backbone, etc.) and you simply don't have enough...

...particular surface receptor are stained with a fluorescent antibody against the receptor and analyzed on ...fluorochrome is removed from all other detectors except its dedicated detector. To ensure proper compensation ...directions. The degree of scatter is captured by detectors and provides useful information about the cell...emits a fluorescent signal that is captured by detectors. The strength of the emitted photons varies from...to further separate the B cell population into progenitor B cells, immature B cells, plasma cells or others...flow cytometry experiment depends on a variety of factors including sample preparation, staining procedures...

...as the identification of additional interacting factors. Problems with Existing Methods for Studying Protein-Protein...PPI identification techniques rely on protein interactors to stay in close contact with the bait protein...for providing a global overview of potential interactors with a bait protein but the flip side is that...When this interaction occurs, BirA* activity is restored, and nearby proteins are labeled with biotin. ...lysed, protein extracted, and labeled protein interactors purified via streptavidin pulldown. Following...Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally defined protein...

...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...in education advocacy non-profits like Phipps Conservatory or the Brilliant Club and share your knowledge... photo competition. Or nominate yourself as an educator for a short, animated TED-Ed lesson. 6) Network... pros Where do you meet like-minded science communicators? Attend the annual flagship meetings of AAAS...grow your network. Get to know and get known by editors and others whose careers are in advance of yours...or upload your data to Zooniverse. The Natural History Museum has downloadable resources to enable you...bananas as a Marie Curie Fellow at The Sainsbury Laboratory. She is passionate about science communications...

...), Antibodies: a Laboratory Manual, 2nd Edition. 2104. Cold Spring Harbor Laboratory Publications, New...born before 1985, you might remember going to the store and buying CDs when you wanted to hear a piece of...technology makes it easy for scientists to share, remix, store, and track these essential components of the scientific...production and subsequent purification.  Academic laboratories and companies have already begun creating plasmids...reproducibility. Non-recombinant antibodies are notorious for being the source of irreproducible data (Begley...useful in perpetuity.  Second, plasmids are easy to store and share, making recombinant antibodies the most...

...Hands-off super busy types, who never have time to mentor you, and Micromanagers, who question every detail...the needs of other people. Some students and postdoctoral fellows are hesitant about expressing their ... 5) Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor...Look at the problem from their viewpoint, and brainstorm about solutions that will meet both of your needs...other supervisors might be nice people, but poor mentors (super-agreeable friendly types, indecisives, and...graduate students since 2009. Dora is also the creator of the “Finish Your Thesis” online course.  To ... Read Joanne Kamen's Post on What Makes a Good Mentor Resources on the Addgene Wesbite Find plasmids...

... by guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Golden gate assembly...restriction enzyme specificities at each step. Factors such as enzyme efficiency, stability, and buffer...four base overhangs after cutting. The destination vector receives the assembly after similar Type IIS restriction... inserts. After ligation of the insert into the vector, the BsaI site is no longer present, allowing multiple...the original insert religates into its original vector, it will subsequently be re-digested and "eligible...our guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Becky Kucera is...assembly, New England Biolabs, Inc.         Eric Cantor is a molecular biologist and a Development Group...

...Gateway pDEST, and pBAD vectors for example. Protein expression from each of these vectors is controlled by ...which expression system to use depends on several factors: The protein you are trying to express How much... interest is inserted into a plasmid expression vector that is then transformed into a bacterial cell....cellular debris. There are several popular DNA vectors that can be used to produce large amounts of protein...resulting in different levels of expression from each vector; lower expression may be required if your protein...protein is toxic to E. coli. Of all the vectors, pET, under the control of the T7 lac promoter and induced...

...with a Cas3 effector, type III with a Cas10 effector, and type IV with an unknown effector. Type I has...repertoire of CRISPR has brought many other Cas effectors to light. One lesser-known Cas is waiting for...genomic alterations on a scale that other Cas effectors can’t, while maintaining relatively high efficiency... family, which means it requires multiple Cas effectors to carry out genomic editing. Specifically, these...still be a challenge. A multitasking Cas Most Cas effectors have one job — Cas see DNA, Cas chop DNA. Cas3...

...Zhang Lab - Multiplexed Overexpression of Regulatory Factors (MORF) Library 3,548 250 - 10,000 bp,...more difficult to transform.    Figure 1: Factors to consider when creating a pooled library amplification...Created with BioRender.com.   The most important factor to consider is your desired library coverage, since... type of cells you are using, as well as other factors, such as your lab’s normal transformation protocol...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...inserts of different sizes (such as a transcription factor overexpression library) or is prone to recombination...

...in 2004, we were the plasmid repository. In 2016, we added viral vectors, and in 2022 we added ready-to-use...ready-to-use antibodies. Now, we think of ourselves as a repository for DNA-based materials, including our popular... kits and libraries. This DNA-based materials repository contains 139,868 reagents from 5,821 labs in ...international community, we've partnered with distributors in China, Korea, Japan, India, and Brazil.  ... system for scientists using materials in our repository, from our customer and science support to our...

...amazing stories to share. I spend a lot of time telling scientists about Addgene’s repository, and explaining...student, post-doctoral fellow, lab manager, research technician, or Principal Investigator. How-to guides...for genome editing? Know a professor with a great story to tell? Let us know by emailing us at [email protected] on Twitter and Facebook. We’ll track down the story, get the inside scoop, and keep you informed.   ...

.... (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs ... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...expression system Choosing an appropriate expression vector is the first step in generating transgenic Arabidopsis...from the Tsuyoshi Nakagawa lab are binary gateway vectors and can be used for promoter swapping and generating... ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology’, Frontiers in Plant...

...m a father of three, a scientist and science communicator, and I strive to be a volunteer and contribute...scientist. When our elementary school needed a new coordinator for the annual Math & Science Night, I took the...contacting our school district’s science curriculum coordinator with minimal luck, but eventually the 8th grade...for a scientist. I think that establishing this history helped demonstrate that a) I was a real person ...great feedback from teachers and other science communicators.  What can CRISPR do? I spent more time on...which demonstrated to the students that scientific history was very much in the making. One of the exciting... in an exciting, tumultuous time in scientific history. Whatever your reasons, let us know how it goes...

...significant holidays. Figure 4: Plasmid request trajectories suggest an exponential decay demand model with... scientists or labs placing orders.  Our model factors the likelihood in our Poisson process into two ... multiplies the likelihood by the corresponding factor 𝜙z if t is during event z. The "events" we considered...each region using stochastic gradient descent in Pytorch, which gave us the maximum likelihood estimates...chemistry and chemical biology.       David R. Liu is Director of the Merkin Institute and Vice-Chair of the ...University; and Howard Hughes Medical Institute Investigator. Liu’s research integrates chemistry and evolution...four examples of technologies pioneered in his laboratory. Learn more here. Additional resources on the...

...proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal circuits. Epitope ... is therefore difficult to use epitope tags to monitor single molecules - something researchers often ...tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster - A... CA3 pyramidal neurons which are known to be notoriously difficult to label. It has also been shown that...technologies such as array tomography, super-resolution STORM imaging, and electron microscopy. Sample preparation...standard proteins tags like GFP. Specifically for STORM, the high intensity signal provided by smFPs can...Spaghetti Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV preps ...

...Rogers is a postdoctoral researcher in Patrick Müller’s group at the FriedrichMiescher Laboratory of the Max... post was contributed by Katherine Rogers, a postdoctoral researcher at the Friedrich Miescher Lab of ... response to an experimental treatment can be monitored. This can reveal previously unknown functions ...splicing. (RNAi, in contrast, engages existing gene regulatory mechanisms to block activity and does not work...phiC31. The popular UAS/Gal4 and Cre/Lox gene regulatory systems have also been successfully implemented..."Single-cell reconstruction of developmental trajectories during zebrafish embryogenesis." Science 360.6392...