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... articles. Here's what you'll find in this post: Glutamate indicators with increased sensitivity and...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation... for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean,..., Iyer, R., … Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation...and AAV plasmid sets launching later this year!  Find Addgene's own Lentiviral backbone plasmids here!... 5% and 11%. It has higher efficiency and lower indel formation than methods relying on homology-directed...scarless, unlike previous integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger, L...

...options critically. How will it feel when you find it? If you find the right lab, some days it won’t even feel...scientists seem to toss data-driven reasoning out the window when making decisions. We think scientists make...success. Gather data on the lab choices How do you find out what the lab is really like? Gather data of ...boss You sign on to a lab for the mentoring only to find out that the lab head is an absentee advisor. They...character traits and skills that you hope to grow.  Find mentors who take their responsibility to train you...meaningless tasks, removal of responsibility, repeated reminders of blunders, setting target up to fail, shifting...(topic and/or technical aspects) applicable to industry if this path interests you  (e.g. human disease...

...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...deposit please sign up here.  Here's what you'll find in this post: Tools for labeling excitatory inhibitory...respectively. Once expressed in the neuron, the FingRs bind to the endogenous synaptic proteins allowing them...inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration ... show a high proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in ...in high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...a few highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene...

...at Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET... particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation...directly interact with the bait from those that indirectly interact or from proteins that are just in the...labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein and saturation...Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting proteins... PMID: 28585547. PubMed Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann...fragmented at a different site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim...

...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find new CRISPR tools for your research! This time: ...control. Read the paper in Nature Communications Find the E. coli genome-wide CRISPR inhibition library...fold. Read the paper in Communications Biology Find the Cas9-PCV and PCV-Cas9 plasmids Selective curbing... editing efficiency. Read the paper in Nature Find the SECURE base editors All-in-one AAV delivery... in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR interference...essential C. albicans gene. Read the paper in mSphere Find the dCas9 and dCas9-Mxi1 plasmids If you have a...

...most prepared of us can find ourselves talking to our posters, so if you find yourself in this situation...session, or it may be held at an unfamiliar venue. Finding your poster board well ahead of time ensures you...at the actual venue. Introduce yourself: I always find it odd when I walk up to someone’s poster, and then... a lap around the posters and read the titles to find the ones you’d like to revisit. Strategically traverse...give the “elevator pitch,” of his/her project – kindly let the presenter know you are very interested ...either seek out the presenter after the session or find him/her on social media or through their affiliations...who presented on a topic related to your research? Find a way to get in contact with the presenter after...

... weak gRNAs (CFD < 20) bind almost not at all, and strong gRNAs (CFD > 80) bind almost as much as perfect... when designing gRNAs Find other CRISPR software Resources on Addgene.org Find CRISPR plasmids and resources...rest of this article, we'll explain the thinking behind the tool, its key features, how it works, and how...we began using it internally. Guiding principles behind CrispyCrunch While CrispyCrunch may evolve over... not available publicly. Furthermore, we did not find any tools which were optimized for HDR, except for...typically advised to mutate the PAM to which the guide binds. However, if the PAM resides in a coding region,...single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR may inadvertently...

...specific epitope that your antibody binds. For example, Myc1-9E10 binds EQKLISEEDL (pronounced “equi-kli-...Resources on Addgene.org  Find recombinant anti-c-Myc [9E10] at Addgene Find plasmids with the myc-tag...same antibody developed almost 40 years ago? Well…kinda. In some cases, antibody providers are distributing...to protein engineering. As a result, you can now find Myc1-9E10 in a variety of isotypes like rat IgG1...this trusty tool doesn’t end there: you can also find anti-Myc [9E10] Fab fragments and single-chain variable...accessibility and the variety of formats you can find it in likely mean it will remain an important tool...tool for the foreseeable future. These newer findings simply add to our understanding of how this antibody...

...survey. When asked if they could detect indels (an indication of gene disruption ability) with NgAgo, ...surveyed here could not achieve indels with the NgAgo system indicates that it is very tricky to use and...many researchers were having difficulty achieving indel formation (gene disruption) with this new technology..., one respondent out of 88 confirmed successful indel formation at a target site. Also, 41 respondents...dead in the water. On the bright side, successful indel generation and epitope knockin was achieved by a... a few respondents suggesting that NgAgo can indeed be optimized for genome editing in mammalian cells...transfections with the ssDNA guide, although many may find this inconvenient. Another possible reason for the...

...the transposase enzyme binds the transposon’s long terminal repeats (LTRs), induces a double-stranded break...A DNA footprint is left behind. When the transposon-transposase complex finds its target site, it integrates...surveys the genome and binds its target DNA sequence. The transposase complex binds the mini-transposon,...for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from its terminal sequences...and unlike other transposons, it does not leave behind a DNA footprint sequence after excision. piggyBac...Transposons are also less likely than viral vectors to induce an immune response and are also easier and cheaper...

...interference, can reversibly exert their effect independent of gene copy number. RNA interference (RNAi) ... 1). As a result, gene silencing can be easily induced by exogenous application of long or short dsRNA...applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi...experiments that require exogenous protein, successful induction of RNAi only requires the presence of dsRNA. In...phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long hairpin RNA.... Each construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene...more specific than long hpRNAs, they must be individually engineered and empirically evaluated for their...

...Antibodies Resources on Addgene.org Find recombinant antibodies at Addgene Find plasmids encoding recombinant...targets, preventing your primary antibody from binding (Fig.1). The level of cross-linking that occurs...temperature, so you may be able to recover good antibody binding by tweaking those parameters, but in some cases...So, while organic solvents may improve antibody binding for some targets, they can destroy it for others... 1: Choice of fixation method impacts antibody binding. Formaldehyde cross-links the protein (red lines...results by not permeabilizing as you’ll prevent binding to any target protein that is still on the inside...reveal otherwise hidden epitopes. However, keep in mind that you will still contend with many of the same...

...through winning and losing funding, and by finding our individual niches within the field of grants. What...want to be an independent grant consultant, building your area of expertise will help you find work and target...serve time and resources. 3. Get a mentor, find your people Find someone who is working or has worked as ...career as a Grant Professional has been a long and winding path for many in the field. The vast majority of...completed. To enter and advance the profession, you can find professional development, certification, degrees...career. Do you have a passion for the arts? Do you find satisfaction in helping your local food shelf raise...experience and to build upon your existing skills.  2. Find your niche Identify and develop an area of expertise...

...their work (H-index for instance), but these rely upon other researchers being able to find their publications...that another researcher finds useful, that researcher will be able to easily find the fruits of your labor...depositors, Addgene strives to make it easier for you to find the plasmids and publications relevant to your research...only does citing plasmids from Addgene help others find them later if they need to use them in their own...similar projects and should make it easier for them to find your work later. We also hope that this feature ...instance, on our Optogenetics Collection page, you can find tables containing plasmids for expressing different...Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

...-throughput sequencing to identify genome-wide binding patterns of the target protein. Both IP/MS and...bottleneck, however, to these IP-based assays is to find (or generate in-house) a pre-validated antibody ...tag-specific antibody (Jarvik and Telmer 1998). Indeed, this method can be used at high throughput because... transcription factors in a short (3-week) time window; this was possible because reagents could be purchased...Ensembl genome browser: https://www.ensembl.org/index.html Design crRNA/ssODN: Using Ensembl Transcript ...recent article by Clarke et al shows that crRNAs binding to the template strand (strand used by RNA polymerase...knock-in efficiency CRISPR reagents had crRNAs that bind to the template strand. Therefore, I suggest that...

...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...the endogenous enzyme is not efficient enough to hinder adenine base editing. Once base transversion editors...C-terminal domain of Cas9, which narrowed the editing window and reduced A → G bystander edits. Their final ...these adenine transversion editors produce a lot of indels (up to 40% at one site tested with AYBEv3!). Although...substrate for a base excision repair pathway. The team behind AYBEv3 introduced the first deaminase-free base...transversion editors, this editor had relatively high indel rates ranging from 4.7% to 30% at tested sites. ...deaminase-free C → G editor with a different editing window than existing CGBEs. This distinct approach means...

...distributors (Table 1), in China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor... Mexico e-nnovation Life Sciences 2020 India Sinapse Biotecnologia 2022 Brazil   ... large percentage of these shipments arrived in India at room temperature. Partnering with our distributor... items to countries with a distributor. Arrows indicate the year we entered into an agreement with a distributor...accessibility to our repository by making it easier to find materials and learn about them. As early as 2007...packaging systems, and luciferase expression. On individual material pages, we added curation fields for ...Collections Lentiviral Plasmids Tetracyline Inducible Plasmids Empty Backbones Bacterial Expression ...

...Development and Discovery As part of their quest to find, characterize, and develop new CRISPR systems and...tests samples of new bacteria from anywhere they can find, whether the samples are from a rain gutter or the...lab for testing, they weren’t sure what they would find… but it was safe to say the sample had their attention...inhibition when the system was retargeted. This finding was surprising – in the most common RNA targeting...further experiments strongly supported their initial findings.  The team originally suspected that this new ... any gene editing treatment would have to be individually customized to the patient's specific mutations... via Site-specific Targeting Elements (PASTE). Finding the right integrase for this new approach proved...

...   Thinking Outside the Box: Individual Approaches Whether it’s finding the source of contamination or...innovation. If you identify as a non-creative or if you find yourself in a creative rut, read on!   Fig... a colleague. To innovate, you’ll have to focus. Find yourself a block of time and a quiet space where...When they were instructed to use the three-ring binder as a starting point, the ideas generated were hardly...get around the lab faster that way— but the idea behind the study holds. To generate novel ideas, start...Team Approaches At some point or another, you’ll find yourself needing to brainstorm with a team. One ...t had time to process on their own. You may also find your meetings run smoother when people have thought...

...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...machinery proteins, or directly inhibit replication by binding to the origin of replication and blocking replication...the first, the replication initiation factor RepA binds to the iteron to initiation replication. However...However, in iteron-containing plasmids,  RepA can also bind to its own promoter and inhibit its own transcription...handcuff” model, iterons of two separate plasmids bind to the same RepA molecules, forming a “handcuff”...region on the plasmid, a centromere-like region binding protein (the CBP), and a partition NTPase (Schumacher...Schumacher, 2012).  During cell division, CBPs bind to the centromere-like region on each plasmid and pair...