... voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...found an indicator that is optimized for 2-photon imaging of voltages. This includes monitoring of fast... plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think we missed...what you'll find in this post: Improved voltage indicator: JEDI-2P Novel class of light-gated potassium...successfully developed an improved voltage indicator (GEVI) called JEDI-2P. Using a custom, high-throughput... available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and you can also find several other constructs...screen for improved genetically-encoded voltage indicators (GEVIs) based on the ASAP sensors. The resultant...

... to deliver larger cargo such as base editors and prime editors. While using AAV to deliver large transgenes...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of...feasible, it leaves little room for additional regulatory elements. Feng Zhang's group previously packaged...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...for Cas9. However, to allow for more room for regulatory sequences while still fitting Cas9 and its gRNAs...Split inteins have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW...

...including the lung, the heart, and the abdomen. Long story short, when asbestos is breathed in (or less frequently... for years, or they are exposed even one time. Doctors and researchers don’t entirely understand why some...do, but we do know that there are some genetic factors involved. For example, multiple studies show that... being done to see what, if any, other genetic factors may be in play. However, there is no question that...asbestos exposure remains the single-most important factor in developing mesothelioma. Asbestos, and certain...Early detection, immunotherapy, and angiogenesis inhibitors New research is also going into two other areas...exposed to asbestos Tests like these will allow doctors not only to detect mesothelioma much earlier, thus...

... Daniel Mejías Sánchez, PhD, traductor  L. Denise Roche, PhD, MD, editora, [email protected] The ...with distributors across the world to help increase global access to materials in our repository. This...  Daniel Mejías Sánchez, PhD, translator  Denise Roche, PhD, MD, editor, [email protected] la traducción, Addgene contrató a una traductora científica profesional, L. Denise Roche, para...el acceso global a los materiales de nuestro repositorio. Este increíble regalo permitirá que más personas...Addgene contracted with a professional scientific translator, L. Denise Roche, to edit the translation. We...

...skeptical reader of historical fiction (I can personally attest to this!). Following history from the Ice Age...relevant science."     - Randall Carter, Contracts Administrator Inferior: How Science Got Women Wrong - and ...and the New Research That's Rewriting the Story by Angela Saini "This book is a smart and carefully presented...science of race."     - Joanne Kamens, Executive Director  I Contain Multitudes by Ed Yong "Microbes are... Tsang, Science Communications and Marketing Coordinator Non-science favorites  The Mistborn series by...another secret."     - Marcy Patrick, Associate Director of Quality Control Jonathan Strange & Mr. Norrell...Norrell by Susanna Clarke "A quasi-historical/fantasy fiction. Two dueling magicians in the 1800s in England...

...you can usually see a director sitting in their chair not watching their actors live on set, but peering...about getting in close to mic them up. Monitor: I then set up a monitor connected to the wireless transmitter...camera, OR, monitor from that distance.  Build: We got a wheeled stand and mounted a monitor to it. It ... their mask. But while the camera is here, the operator is in a different room.   Design: The previous...peering at a 7-inch wireless monitor. Wireless video technology has gotten very impressive, but due to costs...the beating heart of Addgene. It’s why we exist, storing plasmids, and distributing them to labs that need...Cycle 5 - The Future! Figure 5: The mobile monitor and teleprompter being repurposed for a different...

... addition, there was an entire story-telling session organized by Story Collider and which featured a ...new knockout mice, using adenoviral vectors to deliver IGF receptors to the brain, and probing how fatty...to make efficiently. For select viral vectors in the repository, you can instead have Addgene make the...the different parts of the brain stimulated by stories. In retrospect, I should have expected this given...helmets, gait tracking technologies, and brain monitors. Walking around the poster presentation area, ...available from Addgene. While super useful, these vectors require a bit of expertise and can be difficult... into neuroscience, you could look to our viral vector pages on the Addgene website and on the Addgene...

... 1. Testing kinase inhibitors Zhou et al examined the ability of kinase inhibitors to alter Raf1-MEK-ERK...kinase inhibitors, psMEK1 and ERK KTR-mRuby2 expressing cells were incubated with an inhibitor of choice...signaling pathways, scientists frequently use growth factors or serum to stimulate a pathway of interest, but...endogenous ERK, and its localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates...localization of ERK KTR-mRuby2 determined. If the inhibitor targeted MEK1 or its downstream target ERK, the...the KTR remained in the nucleus. But if the inhibitor targeted upstream members of the pathway (Raf1, ..., which frequently occurs when serum or growth factors are used as a stimulus. This system also allows...

...one 30-minute reaction. The destination vector and entry vector(s) are placed in a single tube containing... appear in the final construct. The destination vector contains sites with complementary overhangs that...TGGA and 3’ overhang TCCG can be ligated into a vector containing those overhangs. Entry DNA overhangs...enzyme and ligase. Although the original destination vector + insert may spontaneously religate, this transient...for multiple DNA components (promoters, genes, terminators, etc) to be assembled in the correct order in...assembly of multiple DNA fragments to create TAL effector nucleases. These plasmids were designed to utilize...efficiently clone up to 7 gRNAs into one destination vector, making multiplexing easy.  Disadvantages of Golden...

...  Expanding our viral vector service Addgene has offered in-stock viral vectors since 2016, with the goal...Packaged on Request, an expansion of our viral vectors service, has officially launched! As of today, ...3,000 eligible AAV cargo plasmids in the Addgene repository to be packaged in your choice of five different...easy access to high quality, commonly used viral vectors. Of course, many scientists also need easy access...access to high quality, not so commonly used viral vectors. That’s where AAV Packaged on Request comes in.... making it easier to go from selection to viral vector. To start the AAV packaged on request process, ...for you to dig into. Check our blog post on viral vector production and quality control, our AAV Packaged...

...Susanna Bachle, Addgene the nonprofit plasmid repository 49:45 0 1:18:45 - Reagent Development - Steven...Alex Tucker (Ginkgo Bioworks) Pamela Hines (Senior Editor at Science) Edward J. Hall (Professor of Philosophy...sequenced and quality controlled plasmids and viral vectors. When working with a plasmid from Addgene, you ...negative” data We all want to be able to tell a good story when presenting scientific findings. This makes ...of information and, from a SciComm perspective, stories with links of causation from initial hypothesis...validation are easier to understand. The need for great stories has, however, made it difficult to publish so-called...original hypotheses. Looking for novelty in our stories, we are additionally averse to producing papers...

...industry, non-profit, and government sectors.  Within these sectors, you could start your own company, ...based on any or all the contents of this post.  An attorney should be contacted for advice on specific legal... as law, medical/technical writing, clinical, regulatory, product development, business development, consulting...career paths in academia as well as the non-profit sector are transferrable to industry such as technology...greater scientific field, potential customers, investors, or other relevant entities or individuals. A ...peer reviewer for journals, acting as a journal editor) of the work of those in your field. Original contributions...papers that you should start contacting journal editors to request to be a reviewer. Proactivity is very...

...for drafting my stories. In total, I prepare 5-6 stories for an interview, since one story usually has more...Instead, I preferred to prepare stories to answer interview questions. Story telling shares many of the same...of the theme of my interview stories as the characteristic which the story demonstrates I possess.   Bullet...helpful for my interviews in industry. Prepare stories to tell in your interview The STAR interview method...answer: a setting, a conflict, and a resolution. Stories usually have a theme too, which is something not...of questions. Writing a rough outline of these stories helped me remember them for the interview.  Prepare...

...including a handful of plasmids that have been in the repository for many years, has now grown into a collection... spike protein to bind ACE2, the host cellular receptor. To more easily study the spike variants, scientists...Choe’s lab identified that ACE2 is the functional receptor for SARS-CoV. The lab also found that cathepsins...have an important role in viral entry, which are inhibitors of SARS-CoV-2 entry and become upregulated during...pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames and identified...proteins which all but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-...above, some of these plasmids have been in the repository for many years while many were developed during...

...time: Functional genomics research in an introductory laboratory course. CBE-Life Sci. Educ. 17, ar1. (2018...experiences to undergraduate students (1, 2), but educators face significant challenges in designing suitable...could provide the basis for student projects in introductory labs. Genome projects use computational methods...research questions. The idea is straightforward. Instructors first select a multi-gene pathway to study and...yeasts are thought to have diverged from a common ancestor ~350 million years ago. Over time, the two genomes...species into the pYES2.1/V5-His yeast overexpression vector. This plasmid contains origins of replication and...research skills (8). A complete set of student and instructor resources is available at our Investigations ...

...proteins for use as FLIM-FRET acceptors  Listen to the FLIM-FRET acceptor podcast segment Article contributed...FLIM-FRET acceptors here. See Addgene's FRET reference page for many other common FRET vectors. Murakoshi...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...termied soCoChR, by fusing a fragment of the kainate receptor subunit to the C-terminus of the protein. Unlike...soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish real and artifactual postsynaptic...placesTagRFP-T and Dronpa in close proximity. By monitoring changes in TagRFP-T fluorescence intensity at...Murakoshi lab has developed two new FLIM-FRET acceptors, ShadowG, a novel dark green fluorescent protein...

...primers that target the common features of viral vector backbones; once validated these universal primers...higher than for functional assays depending on the vector backbone. When using qPCR based methods for viral...plasmid which can over-inflate results. A packing vector deficient in reverse transcriptase will produce...the number of target cells that are positive for vector encoded transgene expression following transduction... prep. This method is particularly useful when vectors carrying fluorescent markers are used as it eliminates...significantly higher than the same prep that has been stored in the freezer. If you plan to freeze a batch of...titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009): 495-505. PubMed PMID...

...We’ve dug into the data from our repository to find our most requested plasmid in 2015. In line with ...that the plasmid most requested from the Addgene repository in 2015 was... lentiCRISPR V2   LentiCRISPR...fantastic research tool has been requested from the repository nearly 1,000 times in 2015! If you haven’t already...suggests, lentiCRISPR v2 is a lentiviral transfer vector used to deliver both Cas9 and your gRNA of interest...lentiviral titer as compared to previous lentiviral vectors. It can be used both to knock-out single genes ...is available from Addgene both as a single empty vector and in pooled libraries containing thousands of... has already been used for includes work probing mTOR signaling in cancer cells, deciphering the mechanism...

...recently we have also been using Addgene CRISPR vectors to generate cell lines with knockouts of specific...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings ...sterile truncated pipet tips. Indeed, an experienced operator can pick 48 clones from a 10 cm dish in 15- 20...medium containing serum in each well. A 200 ul pipettor is set to 200 ul, wiped down with 70% ethanol,...individually punched out, gently sucked up into the pipettor, placed into wells, and partially dislodged from...YouTube video. Before returning the plate to the incubator, additional growth medium is added to about halfway.... We highly recommend the use of an electronic pipettor with variable spacing (such as the Viaflo from...

...variant can then be fused to effector domains such as transcriptional activators or repressors. These fusion...cut either strand of DNA. Traditional Cas9 regulators In addition to its utility in making targeted...to DNA, Cas9 can be reprogrammed to serve as a regulator of gene expression. Its catalytic domains can ...hairpin, that is capable of recruiting additional effector domains (4). Taken together, these minor gRNA ... generate a potent Cas9-based transcriptional regulator without making any functional changes to Cas9....adapter capable of recruiting a transcriptional activator, robust regulation of gene expression is observed...the truncated gRNA recruits a transcriptional activator to the Cas9-gRNA complex. When this complex is...