...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...

...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...

...), 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...

...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...

...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...

...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...

... 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...

...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.   ...

...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...

...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...

.... (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...