...plasmid DNA transiently (similar to bacteria) or incorporate the genetic material directly into the genome...not used for general plasmid selection. Instead, reporter genes are often employed to easily monitor transfection...expression levels in the cells. Ideally, the chosen reporter is unique to the cell, is expressed from the plasmid...assess transfection success. GFP is often used as a reporter and we will be covering this ever-versatile fluorophore...population of cells that have fully and successfully incorporated foreign genetic material into their genomes....should be a selective pressure to force cells to incorporate the plasmid DNA into the genome. For the purposes...Before doing a transfection experiment, it is important to determine the proper concentration required...

...antibody isotypes important? To your immune system, an antibody’s isotype is important because it determines...cells that express receptors that recognize the Fc portion of different antibody isotypes. Binding to complement...that helps attack extracellular pathogens.  Transportation of antibodies to places they can’t reach on...region, can be bound by special receptors that transport antibodies through cells and into different body...resources References Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune System in Health...www.ncbi.nlm.nih.gov/books/NBK27106/  Janeway CA Jr, Travers P, Walport M, et al. Immunobiology: The Immune System in Health...

...lab. Granted, I currently work on the Scientific Support Team here at Addgene, so troubleshooting molecular...thesis as it is for helping your team reach an important milestone at work.   While I was in graduate school...problem solving skills’? As a part of the Scientific Support team, much of my day is spent helping to resolve...problem, you might even be expected to write up a report to communicate your findings to your colleagues...work through it. Ask yourself if you missed any important details. Or challenge yourself to think of more...strengths! Research experience presents ample opportunities to practice problem solving skills and learn...that this skillset is a part of your toolkit is important. Paired with your other skills, both technical...

...arrived in 2012, our repository was rapidly able to support sharing of CRISPR constructs and information across...AAV vectors, as well as developing and sharing supportive educational resources. By 2017, we had distributed... the COVID-19 pandemic, and Addgene pivoted to support researchers combatting the virus. Our COVID-19 ...Addgene began to think about how we could expand our support for the research community once more. Our ready-to-use...another plasmid-based product that, with Addgene’s support, could fill a community need for high-quality service...Neuroscience AntiBody Open Resource (NABOR) platform, supported by an NIH BRAIN Initiative. We also created a .... And we look forward to twenty more years of supporting you in your work, your research, and your discoveries...

...vast network and ability to identify the right opportunities, he was able to bring together talented and ...impact. When she spoke with Teytelman about an opportunity to work with a collaborative group, she also ...connection with Joanne Kamens, Addgene also had the opportunity to contribute to this open science collaboration...was particularly helpful for securing funding to support this effort. The future of reproducibility workshops...Science mini-grant, which will fund a project to support a dedicated website to host all the materials in...network well to seek out valuable and impactful opportunities - There are many brilliant minds out there, ...collaboration is that it is driven by academics, with support of industry partners, not the other way around....

...communicator at the Portuguese science museum "Fábrica Centro Ciência Viva" in Aveiro, Portugal. If cancer was...communicator at the Portuguese science museum "Fábrica Centro Ciência Viva" in Aveiro, Portugal. She is particularly...let’s take a look at exosomes. Proficient in transportation, exosomes are tools of communication and provide...in targeted cells. In this case, exosomes may transport miRNA from drug-resistant cells to sensitive ones..., the microenvironment backstage play just as important of a role if not more.  Many thanks to our guest...

...dietary origin) that are diffused or actively transported from cells, traverse the body through the bloodstream...comprehensive datasets that collectively address many important and fundamental questions in nuclear receptor ...states, their circadian regulation, as well as in important physiological states such as adipogenesis and ...the development of a vast number expression and reporter plasmids and the free exchange of these vectors...characterization of NRs and coregulators, and although the reporter gene has over the years evolved from chloramphenicol...for a reasonable fee. We at NURSA recognize the importance of this service to the NR research community ...coregulators. We look forward to growing this important relationship with Addgene for the benefit of the...

...Grant writing offers various skill building opportunities that graduate students and postdocs should actively...and potential alternative routes, Provides opportunities to convince others why a project is the best...understand the RFA. The RFA contains all of the important information required for a successful grant application...figure must be clear, concise, meaningful, and supportive of the mission of the application and the funding...motivates the request for application and offers an opportunity to summarize the limitations of current state... to determine the exact value of costs. It is important to be aware of supply costs per year, how many...the point that was being made, take it as an opportunity to make clearer statements on the next round ...

...Calcium is arguably one of the most important metal ions in biology (sodium and potassium are up there..., and an allosteric modulator. Because of its importance, scientists have developed calcium sensors to...GCaMP6 family of sensors are commonly used calcium reporters with different kinetic parameters. There are three...also available. Detection methods Many calcium reporters use a single fluorescent protein (FP), requiring...unwanted variables. Twitch is a FRET-based calcium reporter derived from troponin C. It is engineered with...for in vivo studies) There are several calcium reporters specially designed for the aims and requirements... Sep 20;111(6):1119-31. Rose T, Goltstein PM, Portugues R, Griesbeck O. Putting a finishing touch on GECIs...

...Targeted and specific delivery becomes especially important for studies in neuroscience where specific tissues...neuron need to be targeted. Targeting is also important in the development of therapies when expression...Picking Ligands Picking the right ligand is also important for building a chemogenetic system. Your experiment...cellular response you aim to manipulate. It is important to consider that the same receptor can elicit ...drug that only binds to one receptor, so it is important to fully characterize the compound, or compounds... ligand binding domains. When coupled to the ion pore domain (IPD) of a channel, the PSAM acts as a gating...DREADD-based chemogenetics in mice. Scientific Reports 9: https://doi.org/10.1038/s41598-019-41088-2 Magnus...

...the fiber optics is probably the least difficult portion of optogenetics (though it does require a significant...materials needed for optogenetics. There are several important considerations with fabrication of the fiber optics...your stimulation. The scatter effect is also an important consideration when preparing the implant itself...will run you about $2-3,000/laser. However, an important caveat is that one laser can stimulate only a ...photons, Chr2 will open but will not be given an opportunity to “reset” and become receptive to light once...wirelessly-controlled optogenetic implants have been reported by several groups [4, 5, 6]. These have the obvious... on optogenetics we’ll delve into some of the important considerations when performing optogenetics behavioral...

...show causality in neural circuits with incredible temporal and spatial precision (I’m not going to discuss...and components necessary to run it? And, most importantly, what are the methodological and experimental...Deciding how you want to use optogenetics The most important question to consider is what exactly do you want...cell type of interest is obviously of critical importance to any optogenetic experiment. There are numerous...of North Carolina) that offer his constructs. Importantly, control viruses (not expressing the opsin) are...for successful optical stimulation.  It is also important to note that some promoters are so robust they...[10]. Unfortunately, opsins are not actively transported and must passively diffuse so, depending on the...

...function improperly. 3. Incorporation into a FRET biosensor A FRET biosensor is a reporter that undergoes a ...Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label of cells, there's...FP dimerization. Equally problematic, several reportedly monomeric FPs are not actually monomeric, at ...fusions Transmembrane proteins, i.e. receptors and transporters, are integrated into lipid bilayers. Fusion ...native gels fail to detect EGFP dimerization. Importantly, all of these assays are performed in vitro. ...help you make better fluorescent fusion protein reporters for your studies and greatly lower your chances...

...academic science; there is most likely abundant opportunity to continue such discussions. Beyond the excitement...spent any time in academia has likely had the opportunity to attend and participate in journal clubs. They...Preparing for the first meeting There are many important pieces before that first meeting takes place to... to ensure success. One of the most important is letting people know it’s happening. Aim for 3-4 weeks...technical dive on single papers could be more important.  Any slides, presentations, recordings, and other...yourself in this situation, considering stopping (temporarily or permanently) the recurring meeting or perhaps...be well planned and carefully organized. Most importantly, they should be regularly assessed for continued...

...a fluorescent emission wavelength in the green portion of the visible spectrum (hence the name), which...maintaining fluorescence. The protein structure, first reported in 1996, is an eleven β-sheet-containing “barrel...solvent. This tightly-packed structure explains the importance of the entire GFP protein, which is almost completely...constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a promoter of...a given cell type. This type of transcription reporting was among the earliest uses of GFP. Fluorescence...protein-protein interactions. In this case, two portions of EGFP are fused to the proteins of interest,...micrometastases in nude mice in cancer studies, act as a reporter for DNA double strand break repair, and to label...

...searching the Data Hub's curated collection of reports (which you can filter by serotype, species, or ... significant time and money. For instance, this report shows that anterograde, monosynaptic connections...our AAV Data Hub Challenge recording to see two reports presented by the submitting scientists, learn more...replicates of your experiment* Functional data, if the report describes a functional vector such as a channelrhodopsin...injection site as well as the target region, if the report describes a retrograde serotype An abstract that...experiment clearly and concisely* *required for a DOI. Reports that meet our minimum standards (* above) will ...

...the materials available to many countries, import and export regulations, differing customs processes, ...materials are available to any country that we can export to — so far, we’ve shipped materials from our repository...In China, direct shipments weren’t able to clear import control. Partnering with a distributor allowed ... resources As Addgene built out our scientific support services, we realized that another availability...developed systems to allow for easier access to export-controlled items by streamlining permissions and...expanding our website, we’ve learned about the importance of smaller, systemic changes, like using alt-...

...very likely be on a team that supports you when you need it. The opportunity to be creative and think critically...hands-on lab opportunities. You can get your hands in nitrile gloves again! The opportunities are there ...gratifying challenge. You’re also getting the opportunity to learn their “language”, and in doing so you...tips from the Addgene blog. You’ll have the opportunity to discover and evolve your transferable skills...If you approach the transition embracing the opportunity for growth, you can tap into curiosity that led...

...Neuroscience Community blog to read reports from bloggers at the meeting – my report is currently in the works....I have an important question: How much coffee does it take to fuel more than 30,000 neuroscientists over...community. It has been great to hear all the words of support from scientists using Addgene in their labs. And...scientific community. All of this speaks to the importance of scientific sharing to the neuroscience community... left for you to make the most of this great opportunity – so if you’re running out of energy, then it...

...you could be sharing your research or finding opportunities to bring science to the community.  We touch...relationships to have any possibility of getting those opportunities. Don't wait until you need a job to meet people...Director Career exploration “You can't wait for opportunities to fall in your lap. Sometimes, in order to ...to do the thing you love, you have to create opportunities. If you find yourself asking, "Why doesn't this..., you are in the early stages of creating an opportunity. That's how Addgene was founded. That's how my...misjudging your own abilities and missing out on an opportunity.” - Sophia Cheng, Senior Software Engineer “Searching...