We narrowed to 1,082 results for: TOR

...reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting an electrode and...successful inhibition of hippocampal neurons. Initial monitoring Second, this may sound rudimentary, but before...exciting—and unanticipated—road of discovery. Monitoring behavioral output The appropriate behavioral ...shock that has become associated with a visual or auditory cue. The hippocampus has long been known to be...versatility and standardization of genetic tools in the laboratory mouse, most optogenetics is performed in this...University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the....    References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala to nucleus accumbens...

... Bank (DSHB), directed by David R. Soll, is a repository for monoclonal antibodies, distributing over ...antibodies to be used as a powerful reagent in the laboratory.   The DSHB focuses on monoclonal antibodies,...include those directed against human transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In ...against new cancer biomarkers. We believe that repositories, such as Addgene and the DSHB, play an important... to ensuring affordable access to materials, repositories provide standardized quality-control and technical...to Our Guest Blogger!   David Soll has been the Director of the Developmental Studies Hybridoma Bank (DSHB...

...Mammalian Vectors By Marcy Patrick Will You Be My Mentor? Finding and Asking for Mentoring Support ...through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and it has...more and more scientists offering to share their stories – about their research, their job hunts, and their... Genes By Marcy Patrick Plasmids 101: Yeast Vectors By Marcy Patrick CRISPR-Cas9 FAQs Answered! ...

...Our plasmid repository succeeds because of our amazing community. The trust of our depositors to allow us...focused on our mission – documenting your plasmids, storing them, providing quality control and customer support...more scientists outside of the US now use the repository – about 50% of plasmids are shipped to labs outside...manage our website, and continue to improve the repository. We love being able to provide a useful (and ... us to help store and distribute their plasmids. The partnerships and collaborations with institutions...

...Of course, all of you toiling away in laboratories this holiday season want the work you are doing to...been to publish in journals with a high “impact factor,” a measure that dates back to 1975 and is based... online access on scientific impact Partly, the story has to do with online accessibility. It is now about... recently reported that the most cited work in history is a 1951 description of an assay to determine ...of scientific publications. Ventures like Impact Story seek to find ways to capture scholarly work that... impact factor...

...of new tools came out including the single base editors, Casilio, CombiGEM, and a variety of pooled libraries...top requested plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab.... TLDR: This is a versatile mammalian expression vector that you can use to express both SpCas9 and your...resistance gene as separate peptides. PX459 This is the vector backbone. This particular backbone can be used ...Overall, this is a versatile mammalian expression vector that you can use to express both SpCas9 and your...mammalian cell lines. Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9...

...Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia Research...tribulations of the process, I’ve provided guidance and mentoring to several people that found my insights helpful...It helps to come from the lab of a well-known investigator. If you only have middle author papers or no... potential collaborations, critiques of your postdoctoral research, etc. The second question is whether...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum, optimization...

...oligo when cloning my target spacers into the PX330 vector backbone? A2: There is no need to add the NGG PAM...degree of off-target effects depends on a number of factors, including: how closely homologous the off-target...recombination (HR) Q7: Can you summarize HR template vector design guidelines? A7: A few notes below are considerations...recently is the double nickase system, with PX335 vector, you need to find two guides for each cleavage ... co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100 bp insertions...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example/...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good reference...

...simplifies your viral vector options. Almost any AAV transfer plasmid in our repository can now be packaged...522 organizations in 41 countries enriched our repository with 15,020 new plasmids — a testament to the...shared progress. But numbers tell only part of the story. This year, we made significant strides in making... our collection, such as blogs describing viral vector serotypes, or hundreds of Data Hub reports detailing...innovation.  In response to a demand for more AAV viral vector options, we launched AAV Packaged on Request, a...

...Chapter 6: The SITN Editorial Process -Vini discusses the importance of the editorial process and peer-to-peer...and training the next generation of science communicators. In this chapter we learn about some of their...social context. Episodes are driven by narrative and story telling. Chapter 5: The SITN Blog - Vini and Amy...graduate student at Harvard University, former Co-director of Science in the News, and member of the SITN...her PhD from Harvard University, is a former Co-director of Science in the News, and is a member of the...

...molecules for use in the clinic and thereby restore motor neuron function in spinal muscular atrophy patients...gene results in the degeneration of spinal cord motor neurons. This disease is estimated to affect anywhere...other genes. He will use fluorescent proteins to monitor the localization of these genes, CRISPR to disrupt...Peritoneal Mesothelioma  Noushin Nabavi is a Postdoctoral Fellow at the Vancouver Prostate Center and ...systems should allow Nabavi and coworkers to test inhibitors of a variety of signaling pathways for their ...

...Project - A regulatory toolbox of MiniPromoters to drive selective expression in the brain.... Depositor Collections...., Wasserman, W. W., Simpson, E. M. (2010). A regulatory toolbox of MiniPromoters to drive selective expression...

...Your Insert and Vector by Gel Purification     Now that you’ve cut your insert and vector, unfortunately...unsuccessful 1), in a vector alone plate with many more colonies than the vector + insert plate (unsuccessful..., if you were cloning a gene into an expression vector, you would want the start of the gene to end up... ligation step. 3. Ligate Your Insert into Your Vector In the ligation step, you mix your purified, cut...Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse Our Molecular...

...same page. Ideally, these meetings include all collaborators, from the lab members who are conducting the...interesting results or ask questions to your collaborators. This helps everyone feel included in the project...materials produced from your collaboration is a key factor in maintaining the group buy-in and morale. Especially...frequently the entire group will meet. You and your collaborators should then hold each other accountable to these...and circumstances permit, think about having collaborators spend time with each other in person. One possibility...workshops are another great place to meet up with collaborators. Being able to present data from multiple aspects...

...Mary O’Riordan, and Victor Di Rita, and, last, homework assignments. The instructors are leaders in the...scientists. Over the course of the class, the moderator discussed individual sections of a paper, the ...quizzes and discussion on that week’s topic by the moderators. The assignments, which consisted of reading ...course was breaking up into focus groups with a facilitator. In the focus groups, we were required to critique... of the paper. They should be able to tell your story for you and they should also be easily understood...

...course of the day. His talk, which focused on the history of genome engineering from ZFNs through TALENs ...Doudna, Feng Zhang, and Keith Joung. All Addgene depositors! Addgene was mentioned specifically in Dr. Zhang's...Science is hard. Chris Voigt, another Addgene depositor, discussed some of the many challenges of creating... of the book on my way to the meeting, and its history of code making and code breaking is wonderfully...addition to the poster session was a first for our repository, and I was proud to be responsible for sharing... sure what to expect in terms of interest, but visitors to the poster were impressed to see how the numbers...

... work with a team to get a lot of stuff done.  Factors contributing to job satisfaction Being a good manager...(i.e. enough pay and benefits to live on), the factors that contribute to job satisfaction are a bit surprising...studies overwhelmingly agree that these are the factors that most contribute to job satisfaction: A feeling...Availability of superiors for consultation, advice and brainstorming. Employees must understand the part they play...team miserable.   A good manager keeps these factors in mind at all times when making decisions, assignments...follow the entire series as it is published. Good introductory reference books on managing people: Skills ...

...ideas. What mentors are available for guidance, intelligent discussion, and/or brainstorming sessions to... your interests. We've outlined some important factors to consider and summarize helpful advice from the...Tips to Achieve Success in Academia". Important factors to consider when choosing a research project After...project along the way. Advice from principal investigators Tom Ellis highlights the importance of keeping...patient, being selective, and talking to peers and mentors will help elucidate which project is right for ...

...cytosine base editors to disrupt gene function by targeting start codons, splice acceptor and donor sites...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles... high G+C content genes, as validated through monitoring the expression of fluorescent proteins and oxytetracycline... a) Cells are transfected with three different vectors and flow cytometry is used to sort populations ...CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available...

...CRISPR Resources Learn More about Viral Vectors Browse the Repository  ...101: Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and tired...While the vast majority of CRISPR plasmids in our repository use Cas9 to cut DNA, Addgene Science Communications...generate knockouts, but how does it work? Guest contributors David Wyatt and Dale Ramsden from UNC Chapel...CRISPR/Cas9 and AAV Adeno-associated viral (AAV) vectors are useful for in vivo gene delivery, but have ...

...with CRISPR–Cas nucleases, base editors, transposases and prime editors. Nat Biotechnol 38:824–844 . https...true" %}{% raw %}"Browse Empty gRNA Expression Vectors"{% endraw %}{% end_module_attribute %}{% module_attribute...href":"https://www.addgene.org/crispr/empty-grna-vectors/","type":"EXTERNAL"},"open_in_new_tab":true,"no_follow... are even more exquisite for the so-called base editor Cas9, which makes DNA changes in the absence of...breaks (Rees et al., 2018). For C>T and A>G base editors, the intended edit must be in a 5 - 10 nt window... guest blogger John Doench! John Doench is the Director of R&D in the Genetic Perturbation Platform at...

... or LinkedIn to learn more about their career trajectory. If there’s one thing everyone loves, it’s talking...can manifest via research and data gathering, regulatory efforts, policy implementation, program management...company who decides how they are going pursue regulatory approval for a new product or a well-endowed ...move into science policy. Read broadly about the historical roots of science policy to connect to your why...example proposals and consider applying for their accelerator for some hands-on guidance.   Check out my webinar...careers, there are many ways to enter this career trajectory. One common route of entry is through a science...working in student government, and teaching and mentoring others (see more in Figure 2). Starting early ...

...Research Campus to give researchers the ability to monitor glutamate release from neurons and other brain ...SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate Indicators for Synaptic Imaging However, the first two generations... domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG induction to express soluble protein...is also available in several ready-to-use viral vectors.    References and Resources Additional Resources... 30377363 Aggarwal, Abhi, et al. . “Glutamate indicators with improved activation kinetics and localiation...

...This post was contributed by Matteo Tardelli, a postdoctoral scientist at Weill Cornell Medicine. Discussing...lab meetings with a focus on discussion and brainstorming Though, we might be forced to work from home...of each talk - time for free interaction and brainstorming. This could be achieved by asking each presenter...problem! You can still creatively collaborate and brainstorm! Image from Joseph Mucira.   Implement new...can be used as a break, but can also be used to brainstorm on data in smaller groups, where for instance...blogger Matteo Tardelli! Matteo Tardelli, PhD is Postdoctoral Scientist at Weill Cornell Medicine in NYC and...

...Addgene depositors have made it easier to track translation in two different ways: by monitoring the first...complexes. Recent work has adapted this technique to monitor local translation and determine if proteins are...fusions to their reporter mRNAs. Using translation inhibitors cycloheximide and puromycin, Halstead et al. ...view for constant tracking. You can then directly monitor multiple rounds of translation from this single... Yan et al. used harringtonine, a translation inhibitor that stalls ribosomes after initiation, to calculate...

...expression of the protein. However, expression of receptors may not need to be as high to achieve an observable...titer may not need to be as high when delivering receptor constructs with AAV. In the case of channelrhodopsins...may not be necessary to concentrate this type of vector to such a high titer when looking for effective... possible, use low-bind or siliconized tubes to store samples as it is more difficult for viral particles... advances in recombinant adeno-associated virus vector production." Kidney international 61.1 (2002): ...Where do they come from and what do they mean? AAV Vector Quality Control: Going the Extra Mile with NGS ...

...lentiviral preps of select lentiviral vectors from the repository for you. We perform test infections with...easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert) and the reduced titer associated... your gene of interest with a variety of Cas9 activators available in lentiviral format. Finally, while...lentiviral format Another popular set of tools in the repository are CRISPR pooled gRNA libraries that can be ...Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...

...devices to our own interests with the content we store in them: music, emails, chats, pictures, and apps...culture and include a counter, lab timer, dilution calculator, and a couple of other nice features. This app...sequence for your enzyme of interest. Lastly, the Tm calculator helps you set up an optimal PCR depending on ...information in the app is restricted to the NEB inventory only, the restriction enzyme sequence resource...acids memorized, but the structures are another story. AMINO has been very helpful when I’ve had to explain...apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme Tool by Promega Material...

...contributed by guest blogger Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver.  Most types...enormous lattice of identical subunits. A brief history of R bodies R bodies have been studied for many...bodies do for you? R bodies are robust linear actuators that could be used in a variety of applications...applications, purified R bodies could also perform as actuators in microelectromechanical systems (MEMS) devices...with their environment.   Jessica Polka is a Postdoctoral Research Fellow with Pamela Silver in the department... Biology Posts  Suggest Plasmid Tools for the Repository Resources on Addgene.org Read our Hot Plasmids...

...resources at Addgene. His background in history and specifically the history of RNAi left him a bit skeptical...to the Advisory Board. John Doench is Associate Director of the Genetic Perturbation Platform at the Broad...Listen to the interview here! Doench majored in history during his undergraduate work at Hamilton College...targets and what confers specificity in the miRNA regulatory process. This early research is still relevant...function really…. Addgene’s role as a central repository for what is useful and what was used once and...

...define what type of factor you are looking for and what process or outcome that factor functions in. The...there is removal of known factors (such as a genes) and the loss of those factors is assessed. The two most... committed to the daunting task of finding new factors in your biological system of interest, the so-called...selected growth conditions. For screens, there is no obligatory growth advantage – that is, you are not specifically...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles... cassette provides increased simplicity for combinatorial genetic screens, and this optimized toolkit ...50 mLs) or through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either...by: Rachel Leeson The large size of CRISPR-Cas effectors and fusion proteins, such as Cas9 and Cas12a, ...by the fusion of Cas12f with a transcriptional activator, CasMINI is able to induce high levels of gene...packaged in AAV. A dCasMini-mediated adenine base editor allows for base editing, while gene editing can...

...cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing various...expression using either TEF1 or GAL1 promoters. These vectors are intended for the high level expression and ...spike protein) cloned into a CMV driven expression vector are also available. These contain no tags and are... genes in E. coli and S. cerevisiae expression vectors in late March. The first set of 383 constructs ...status we call “Archived.” This status allows us to store plasmids and make them available to scientists without...

...have perfected processes to curate the data our depositors share and enrich it with sequencing data; as ...categories — catalog, educational resources, and depositors — each with subcategories such as plasmids and...educational resources, and labs and institutions for depositors (see below). Even if the user experience is similar...scoped to plasmids shows filters for expression and vector type, while a search scoped to viral preps shows... match. For example, matches of a name in the depositor field will rank higher than matches of the same...that contains a plasmid with the catalog ID). Depositor suggestions provide shortcuts to matching PI labs...

...this list, that does not necessarily mean the repository does not have it. That said, if you know of a... don’t hesitate to start the process! Addgene depositors who study chromatin The PIs linked below are ...Chromosome Organization. These aren’t the only investigators in the chromatin field who have deposited with...Addgene website. In-exhaustive List of Addgene Depositors Who Study Chromatin Cheryl Arrowsmith James...SNF-like BAF complex to chromatin after T lymphocyte receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek...Bystricky, Ben Martin, and Berkley Gryder. Addgene depositors or not, your time is valuable ...

...textbook, a laboratory kit, and support during the academic year from remote scientist mentors and the BioBuilderClub...effectively contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom...meetings and newsletters, pairs teams with STEM mentors, and provides a nationally recognized platform ...state-of-the-art lab embedded in LabCentral’s community of innovators that opened in August 2017.  This flexible teaching...BioBuilder! Mary Tamer is a Boston-based writer, editor, and consultant whose work is focused on education...

...-introduced breaks are repaired. The following factors are required for NHEJ repair regardless of end ...XRCC4-ligase IV); together with some accessory factors (PAXX, XLF), this complex holds a pair of DNA ends...Accordingly, NHEJ has a vast toolbox of processing factors, including polymerases (Pol μ and Pol λ), nucleases... sequence (“microhomologies”). NHEJ doesn’t obligatorily introduce indels. Given the end structure of...majority of their chromosomes within a day. Another factor expected to impact repair is that the Cas9 protein... loading of Ku and normal NHEJ activity. Other factors can also influence NHEJ activity. For example, ...

...pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit consisting of 834 ...stem cells. Addgene: Tell me about the history. How had investigators done this kind of work before? Musunuru...Unfortunately, it is time consuming and difficult for investigators without a lot of expertise to create zinc finger...and puts it in the hands of pretty much all investigators. Our kit makes it extremely simple to design...

...already run several reproducibility workshops as Director of Scientific Outreach at Code Ocean and through...well received, and then when the team started brainstorming ideas for the next workshop, they realized that...workshop at the conference, but because each contributor made the materials accessible, anyone could present...and were presented at the ASPB meeting by the collaborators who could participate. Despite the group not...visibility of these materials, Sonali Roy, a postdoctoral researcher at the Noble Research Institute, ... Schwessinger for helpful conversations on the history of the group, lessons learned, and to Sonali Roy...

...there are many factors that interact with sex, and it’s important to analyze these factors during an experiment...blogger Londa Schiebinger, PhD, Hinds Professor of History of Science, Stanford University. Sex and gender...experiment. In the case of stem cells, these factors may include cell type, the disease being treated, ...other hormonal, immunological, and environmental factors. How to include sex in experimental design Recognizing...variable in all NIH-funded research. Janine Clayton, Director of the Office of Research on Women’s Health, emphasizes...