We narrowed to 877 results for: TOR

...plasmids are formed. Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs...problems that I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating.... Over time, bacteria containing the recombined vector backbone will also outgrow bacteria that contain...into two smaller plasmids. The ~3.7 kb recombined vector backbone contains the antibiotic resistance gene...samples? Plasmid recombination can affect viral vectors (LTRs and ITRs), plasmids with multiple polyA sites...recommendations from the depositing lab. If your vector is large (>10 kb) or has a low copy number, try...that in Stbl3 bacterial cultures with lentiviral vectors, the flat white colonies contained the full plasmid...

...initial reversal drive cannot restore the exact original sequence, it can restore the original protein-coding...are eliminated by natural selection, gene drives distort inheritance in their favor and consequently can...carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread through the...mistake would necessarily extend far beyond the laboratory and seriously damage public trust in scientists...gene drive is also the easiest: don't use a DNA vector that encodes both Cas9 and sgRNA. What if I want... can spread through populations of transgenic laboratory organisms but not wild organisms. For example...synthetic sequence will only spread in transgenic laboratory populations with that sequence2. Both methods...

... College of Medicine. Her doctoral research was conducted in the laboratory of Erik Lee Snapp and focused...the cytoplasm. That is, unfortunately, some investigators realize too late that they have missed the impact...environment where it will localize [4]. Most investigators are surprised to learn that FPs are often poorly...environments, including the organelles that compose the secretory pathway, endocytic vesicles, mitochondria, lysosomes...residues form inappropriate disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs ...and a sequence intended to localize them to the secretory pathway, a dark pool that is mislocalized to the...misfolded, blocked from proceeding through the secretory pathway, and retained in the ER. FPs have been...

...then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But...RNA-editing Cas13 enzymes have taken the CRISPR world by storm. Like RNA interference, these enzymes can knock ...in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are ...looked for a CRISPR repeat array near a putative effector nuclease. The Cas13d proteins they identified ...identified other advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both ...WYL domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins...Konermann et al.: CasRx is a highly potent, specific editor in mammalian cells Not all Cas13d proteins are ...

...  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David Root lab is...you! We would like to thank all of our depositors and requestors for making their reagents available and...opening our UK office in 2014, to starting our viral vector service in 2016...we’re excited to help scientists...look back at some of the contributions from our depositors. Below are the 15 top requested plasmids over... be used with 2nd or 3rd generation lentiviral vectors and an envelope expressing plasmid, such as our...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver Cas9...your gRNA of interest into mammalian cells. This vector was the top requested plasmid of 2015. What makes...

...therapeutics such as  (Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in immunotherapy...methodology with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor Michel...Professor Karin about alcohol and obesity as driving factors for chronic inflammation which in turn suppresses...self-limiting effects kick in trying to resolve the inflammatory state by inducing immunosuppressive measures...upregulation of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment in turn...tumor and immune cells in real time. Addgene depositor Professor Max Krummel from UCSF showed some impressive...Throughout the conference aspirin and the use of COX inhibitors was discussed and Professor Caetano Reis e Sousa...

...adenine base editor Earlier this year, Keith Joung’s lab developed SECURE cytosine base editors, which decrease... Nature Biotechnology Find the base editors   Single base editor that induces C-to-T and A-to-G mutations...developed a base editor that combines the abilities of C-to-T and A-to-G base editors into one protein...pyogenes Cas9. They called this editor Target-ACE (adenine and cytosine editor). Target-ACE exhibits increased...research! This time: CRISPRi in Burkholderia Base editors CRISPR toolkit for epitope tagging Lentiviral capsid-based...APOBEC1. Now, they’ve engineered SECURE adenine base editor variants using a protein-truncation strategy with...a target site compared to a mixture of two base editor enzymes in mammalian cells. Read the paper in ...

...activation of keratinocytes, which release inflammatory mediators that trigger the activation of dendritic...recruitment of neutrophils. Though there are IL-36 receptor inhibitors in development, there’s still a lot of mysteries...Generalized pustular psoriasis is a chronic autoinflammatory skin disease estimated to affect 0.27 to 4.6...actually get out of keratinocytes and induce their effector functions. It’s an important process that we need...IL-36 cytokines are a potent inducer of an autoinflammatory loop: keratinocytes both respond to and release...when he was studying thoracic aortic disease. “My mentors Dr. Dianna Milewicz, Dr. Carolyn Bondy, and Dr....with TS (Trolle et al., 2016). Using lentiviral vectors from Addgene, the lab will express these candidate...

...point.   What makes a story? A story is a narrative connecting cause and effect. Stories can and should be... or notes. Listen to their stories and narratives (see below “Tell a story”) and note how they build suspense...hear material delivered in the form of a story. Using stories is one of the best ways to make you and ...me and wished I had been more careful. 3. Tell a story We are biologically programmed to listen and remember...presentations engaging and memorable. If you read my story above about the presentation faux pas, you are more... striking images, creative ideas, and engaging stories as you encounter or experience them. Drop content... all graduate students and postdocs form peer mentoring groups to practice giving talks and getting feedback...

...editing assemblies are sensor-actuators, either transcriptional regulators or chromatin-modifying enzymes...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...s chromatin status into account (Fig 1a). The laboratory of Karmella Haynes at Emory recently contributed...contributed a set of plasmids to the repository that will help epigenetic researchers custom design their own...Examples of histone-modifying enzymes and reader-effector proteins. b) Golden Gate assembly (backbones available...with examples of potential synthetic chromatin mediators and probes. Image from Priode et al., 2021. ...potential tools fall into two categories: probes and editors. A probe array may consist of at least two sensor...

...lab has developed AAV vectors to selectively visualize excitatory and inhibitory synapses in the brain...combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory synapses... find in this post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein system...New from the viral service Labeling excitatory and inhibitory synaptic proteins with FingRs By Jason... proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively. Once expressed...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles... cycle in vivo, particularly if the cells are migratory or in deep tissues. To resolve this issue, Shangqin...

...efficiencies of integrating viral vectors. New Sleeping Beauty transposase vectors Even more recently, Eric ...developed two new types of vectors for the SB transposase system - one set of vectors for robust constitutive...transposase as a nonviral vector for CD19-directed chimeric antigen receptor (CAR) therapy. SB is a valuable...of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both introduce...cells’ defense mechanisms. The use of non-viral vectors, like transposons, avoids many, though not all,...use of viruses is also important for social and regulatory reasons. 1.) The transposon consists of a mirrored... expression (pSBbi derivatives) and one set of vectors for inducible expression (pSBtet derivatives). ...

...combinations of promoters, terminators, solubility and purification tags, connectors, cDNAs, and more to find...site-specific recombinases, regulatory elements, developmental regulators, and lots of viral components...workhorse yeast — K. phaffii (formerly known as Pichia pastoris). If your goal is protein isolation, then the ...kit has a focus on infrastructure with shuttle vectors, several delivery systems which can be engineered...engineered into the TUs, and even viral gene silencing vectors. The Plant Parts III kit has a focus on transformation... examine the effects of different promoters, terminators, enhancers, and cross-cassette interactions in...effects of protein degradation on chimeric antigen receptor signaling (Kim et al., 2024). The EMMA Toolkit...

...chair of the postdoctoral engagement committee and active member of the university's Postdoctoral Association...science on April 22, 2017. While experiments (and editors) can move slowly, news about possible broad changes...in the veracity of its work. Alina Chan is a postdoctoral researcher at Harvard Medical School working...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...health depends on it. Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072...to join the public conversation and to tell the stories behind our research.  That's why I marched for ...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...

...about being an editor - that I learned on the job. The day-to-day work of a science editor In my role as...sorts of different editors. Do you like science journalism? Take a look at our news editors. A fondness for...at our multimedia producer and online editor. And, the copyeditors deal with things like word choice, punctuation...contributed by guest writer Pamela J. Hines PhD, Senior Editor at Science Magazine. Although we only walk one ...be like? Then I spotted an opening for being an editor at Science magazine. That one I just could not ...as an editor at Science, my various interests come together as useful and productive, forming a different...science journalists, or policy makers. The term editor covers a wide variety of careers and jobs. At Science...

...might not be a big factor for you. Know who's boss You sign on to a lab for the mentoring only to find out...choose a mentor that sees you only as a paper making machine. Choose one that is a true mentor, supporting...rotation was with a mid-sized lab with an experienced mentor. The lab was very, very quiet. Every time I asked...some of these questions in the sections below. Mentoring relationships can help identify your priorities...not worked for you in the past. For all of the factors I touch on below, the first question to ask yourself...“which ones matter to me?”  and hone in on the factors that will make the most difference for your success...? Will a postdoc or lab manager be your actual mentor? Is there a chance the supervisor will be moving...

...him a more effective motivator (“be like Tony Robbins!” he encourages faculty mentors.)  Collaboration Maximizes...technical and pedagogical resources, offers mentoring for instructors, and provides opportunities for students...in a year-long biochemistry laboratory : Part II – research-based laboratory – a semester-long research...becoming just the fifth repeat Nobel Prize winner in history. Interestingly, he also adds to the disproportionately...this trend as evidence that the best teachers and mentors pass on more than just technical knowledge, shaping...responsibilities left him with limited time for one-on-one mentoring. His solution? Bringing his research into the ...work identifying new genes required for stem and progenitor cell maintenance in planarians in Developmental...

...has a great team of moderators who facilitated scheduling, filtered out inflammatory questions, and even...and the moderators were on top of any comments that were at all trollish. Kudos to the moderators for keeping...trigger phrase these days). Redditors ask serious questions What the moderators did not delete or suppress...incredibly important to find good mentors and incentivize good mentorship: AMA mechanics: Be prepared! The...Addgene Executive director, Joanne Kamens, recently participated in a Reddit AMA (short for “Ask Me Anything...lists of forums on particular topics. Volunteer moderators set the rules for these forums and try to keep...careers, women in STEM, work-life negotiation, and mentorship, we decided to have her participate in an AMA...

...empty control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows... In this context, the empty vector control shows any effect of the vector/backbone itself on gene expression...hypothesis: proper controls! For every experiment, an investigator needs a standard against which the results can...variable you are testing and not some other unintended factor. Control plasmids are used to minimize any effects...planning your experiment includes determining what factors need to be controlled for in order to eliminate...replicate controls.  Transfection controls: empty vector and internal control A transfection control measures...any effects of the transfection itself (i.e., the vector used in transfection, transfection reagent, or ...

...few inventory items offered domestically, but by the end of 2016, we expanded our viral inventory to 25... is a dual-vector system, meaning it is typically used in conjunction with a second vector that delivers...constructs (i.e., using a dual-vector system as opposed to a single-vector system) has several benefits.... lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide...LentiCas9-blast LentiCas9-blast is a Cas9 expression vector developed by Feng Zhang’s lab at the Broad Institute...such as lentiGuide-Puro. The alternative single-vector systems typically express Cas9 and a gRNA from ...compared to separate transductions with a single-vector system). Tips for using this plasmid Cas9 expression...