...the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially...ten colors (see table to the right); in various reports, Sanes and Lichtman have observed from 90-160 distinct...undergone recombination. This default state was disproportionately expressed by a majority of neurons, limiting... 8 colors. The use of AAV provides spatial and temporal control without the need for germline modification...refinements of the Brainbow technique will provide important insights into the complicated physical organization...

... Impact of Non-Elite Journals” found that the proportion of highly cited articles appearing in lower-prestige... to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles in non-elite...publications. (As an interesting aside, Nature recently reported that the most cited work in history is a 1951 ... the best way to measure impact? As that Nature report also noted, papers detailing Nobel Prize-winning... for the impact of research" (2013) Scientific Reports 3, Article number: 1649 doi:10.1038/srep01649 ...

...all the opportunities you can to be communicating about science with others. If opportunities don't exist... not stack the deck in your favor? Look for opportunities to practice transferable skills in ways that...something big like plan a conference Recruit many supporters and mentors that will speak for you (and you ...get real feedback Write whenever you can, find supportive editors so your first drafts are always excellent...

...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...the Q system from a gene cluster in the fungus Neurospora crassa. The QUAS promoter is less leaky than ... transcription from the UAS promoter. This is important because it allows them to be used together for...

...useful for genetic engineering approaches. Most importantly for this technology, however, is that the transposase...should be tested in the cell line of interest. Importantly, by varying these ratios, you can either maximize...obstacle, a positive selection cassette can be incorporated, and used during the differentiation process...homologous recombination, which results in the incorporation of your specific edit into the host genome. ...transposon sequences.  The use of PBx is particularly important to avoid any possible random re-integration from...lines, most likely due to DNA toxicity. More importantly, unlike Cre recombinase, the PBx is highly specific...

...scientists with an MSc or higher apply. It was important to weigh the cost to benefit of studying a Masters.... I felt then, and still feel now, that it is important to take advantage of the fact that science is ...manufacturing practice (GMP) and good lab practice (GLP) (important buzzwords in industry) as well as skills in industry... Queensland in Australia, this was a perfect opportunity to work in a genetics lab. As a UK citizen, I...and to discover for myself what potential job opportunities are actually like day-to-day. It is all too ...permanent position in academia. It is therefore important for young scientists and PhD students to also ...

...means that scientists-in-training rarely get the opportunity to learn about the myriad of diverse, nonacademic...questions on the internet. To make the most of the opportunity, use a written list and take notes if that is...time to ask personal questions about salary, opportunities for advancement, challenges and disappointments...respond to you). You will uncover interesting opportunities by meeting with people working in diverse and...That way you are always setting up your next opportunity to learn. Additional Reading More "Career Advice...

... is resistance to change, public support is weak, government support is “feckless”.  So what can be done...from Harvard Medical School and a long list of supporters from all of the research centers in Boston.  ...for PIs to include in their annual NIH progress reports information on the Individual Development Plans...will come up with to push for change in their opportunities.  Scientists must do science to be happy so ...

...background.” Just last month in Scientific Reports, Verkhusha’s team reported another advance in the application.... Transparency window That’s exactly what they reported doing in a paper that appeared in Nature Biotechnology...they absorb and emit light in the near-infrared portion of the electromagnetic spectrum – the spectral ...proteins, organelles, and living cells in a spatiotemporal manner, which is not possible with conventional...

...piggyBac plasmids    Tol2  Tol2 was the first reported active DNA transposon in vertebrates. It was discovered...mutations. In these screens, the transposons encode reporter genes, mutagenic cassettes, or barcodes. When ...2021). Target site preference and host range are important to consider when selecting a transposon for a ...pronuclei of fertilized eggs, which leads to random incorporation of that sequence into the zygote’s genome, a...unpredictable. Transposons, however, efficiently incorporate into the zygote’s genome after injection into...Sandoval-Villegas N, Nurieva W, Amberger M, Ivics Z (2021) Contemporary Transposon Tools: A Review and Guide through...

...orchestrate the formation of high-functioning-but-temporary teams that have a specific purpose and finite ...year, or a few months, or even a few weeks. The important thing to remember is that every team should have...when I do come across one. Also, remember, one important feature that all projects have in common - and... What does a good team need? Besides that all-important sense of purpose, there are many more things necessary... function is certainly necessary, but equally important is making sure all team members understand what...their responsibilities are. That’s a subtle but important distinction. If I invite someone to be on a project...

... of single-use items, plastic is a substantial portion of lab waste. Despite the fact that researchers... NEB actually started this program in 1976! Millipore Sigma takes back expanded polystyrene boxes for...environment? Well, you could: Collect the NEB and/or Millipore Sigma boxes from your lab or department and send...challenge you to take to action! These days, it’s important for individuals, companies, and universities to...to work on this issue. Be a champion for this important cause! To get more lab sustainability tips, check...

...consider as you make your choice. The first and most important thing you need to know is your expression system...have to make sure that your plasmid has been incorporated into the host organism, usually achieved with...basal expression level, induce expression at an opportune time or go all out with constitutive high level...Some of the more popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. ...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...

...and have always been, a community resource that supports both toolmakers and tool users. We encourage you...Educational Resources As part of our commitment to supporting your research, Addgene is creating an array of...Through our incredible community, including your support, we have enabled scientists to easily share plasmids...Studies Hybridoma Bank, a nonprofit that has been supporting hybridoma sharing within the community for decades...Immunofluorescence Addgene’s Antibody Guide The project is supported by a NIH BRAIN Initiative grant (U24NS119916 )...

...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...the Cre recombinase, are used to spatially and temporally control transgene expression. Lox sites are directional...sites lets researchers control the spatial and temporal activation of a transgene. Figure 2: FLEX...structure when transcribed which aids in nuclear export of mRNA and helps increase the expression of your...polyadenylation or polyA signal aids in the nuclear export of RNA and RNA translation, and promotes RNA transcript...

...scientists who have knowledge of their product portfolio. After all, you have been their target market ...throughout their career?”, "Have they sought out opportunities to expand their skill sets?", “Have they succeeded... interviews Studying for interviews is really important. For my Postdoc position, I tried to read at least... current sales position interview was just as important. The problem was I knew I had a knowledge gap....teaches you about selling products which is an important part of the business and allows you to work with...

...sub-cellular localization signals, plant markers, and reporters (Engler et al., 2014). This kit is a great starter...upon (its ‘basics’ MoClo plant kit buddy). It is important to understand your experimental needs before choosing...into a multi-TU targeting construct (Cas9, sgRNA, reporter, etc.). This allows for the multiplexing of sgRNAs...basic biology of and gene regulation in the opportunistic pathogen Acinetobacter baumannii (Brychcy et...used successfully for protein localization and transport studies in HEK cells, recombinant protein purification...Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media...

...be it an organic dye or FP) is one of the most important steps in the design of an imaging experiment.  ...Goedhart on the Addgene blog – outlining various important considerations when choosing a FP, but much of...valid email address).  One may also simply attach important references that serve to further characterize ...proteins.  For computer-savvy users looking to incorporate FPbase data in their own projects and applications... and elaboration.  Current projects include incorporation of biosensors, protein evolution lineages, 2...

...this year, I jumped at the chance. Here was an opportunity to see Shinya Yamanaka celebrate the tenth anniversary...different fields. This meeting has given me an opportunity to dive a little deeper into the world of stem... most scientists recognize and appreciate the importance of ethics, the topic can often take the back ...conversations but they frequently seem to be of secondary importance (or tertiary, or non-existant). On the very first... the stem cell community for addressing these important questions that the scientific method cannot address...

...ready to support your team and take responsibility if things go wrong. Here are some important criteria...distributed appropriately. If team members have opportunities to develop and stretch then they will be happier...want to know about right away? Provide adequate support. The manager must be available to answer questions...shared document. Update frequently with progress reports, meeting minutes and plan changes. Written documentation...

...expression in the liver after systemic delivery.  Importantly, this is true in both mice and nonhuman primates... a cleavage free manner. The dCas9-SSAP system reports an efficiency of up to 20%, comparable to commonly... For a one kilobase insertion, the researchers reported an editing error rate of less than 0.3%, both ...using mStrawberry, localization peptides, and reporters that drive the expression of the fluorescent protein...response element (TRE) or kinase translocation reporters (KTRs). Find visual barcode plasmids here! Kaufman...

...aspects of my work but also found fulfilment in supporting my team’s development and career growth. This...check-ins, which helped them stay on track and feel supported. Another team member was highly self-motivated... challenges. Experimenting with leadership opportunities before fully stepping into a managerial role...as they encouraged me to explore small-scale opportunities where I could make mistakes and learn from them... served as a great "baby project," offering a supportive environment to build my management skills. Finally...

...function in terms of its regions. But equally important is the way the brain connects across these regions...is connected to itself across these regions is important for understanding how the brain functions. The...associated virus (AAV) shows relatively low retrograde transport efficiency in most serotypes. Some viruses (e....e.g., rabies) naturally exhibit retrograde transport and can therefore perform retrograde tracing, but...requires high titers for efficient retrograde transport. For recombinase-dependent vectors, this can cause...

...it is limited to only 20 bases, or why it’s so important to choose a target sequence with a nearby protospacer-adjacent... lobes laying open like a book (Figure 2). The portion of the CTD that recognizes the PAM is so flexible..., but they improve cutting efficiency and are important for robust activity in cells (Jiang & Doudna, ... guide RNA stabilizing the DNA-binding protein portion of the CTD, ready to check for PAM sequencines....correct positioning of the non-target DNA strand is important for stabilizing the active conformation of the... University of California, San Francisco, with support from National Institutes of Health R01-GM129325...

... expands learning outcomes by emphasizing the importance of critical thinking, persistence, teamwork, ...emerging scientific field, BioBuilder embraces new opportunities for scientific exploration, discovery, and collaboration...with a kickoff meeting in October, BioBuilderClub supports teams through monthly online group meetings and...the BioBuilder textbook, a laboratory kit, and support during the academic year from remote scientist ...on the Addgene Blog Synthetic Biology and the Importance of Assembly Standards Synthetic Photobiology:...

... non-linear dependence on excitation power is important since different fluorescence imaging strategies...requirement for quantitative live-cell imaging, it is important to be able to quantify the photostability of fluorescent... be noted that the photostability measurements reported in the literature are performed in different ways...information can help guide your understanding of reported data. Photobleaching results in a loss of fluorescence.... To avoid changes in excitation power, it is important to perform the experiments on a single day on ...

... since I was still in grad school, I had the opportunity to take a course through the school of journalism...donors such as fundraising materials and progress reports, in addition to articles for the Fred Hutch website...tough assignments, the skills that Susan  finds important for her work include: “Listening. Evaluating evidence...started in science communication: “Find as many opportunities as you can to get your name on things.” Susan...this easier to do while still in school since opportunities, such as writing for a student-led publication...

...As a scientist, perhaps the most important part of your research is communicating the results of the ...the work. Yes, those control experiments are important - but without letting other scientists know the results...Some organizations, including some journals, also support open peer review or curation models where editors...earlier attention and feedback giving authors the opportunity to strengthen the manuscript prior to submission...Along with other open-science organizations, we support scientists in sharing their results and materials...

... how these structures are generated, why it’s important for cells to sense their formation, and how clinicians...in turn lead to genome instability. Thus, it’s important to strike a balance between productive levels ...be delivered or anticipating this response are important considerations when using these reagents. RNA...recognized by RNase H which hydrolyzes the RNA portion, allowing the DNA strands to re-pair. The two dominating...stranded isn’t just for DNA. Untwisting RNA is important for therapeutic delivery, viruses, human disease...

...different ways depending on the team. When Tony, our corporate counsel, used the word “service,” did he mean ...mean it the same way as Bryan, our customer support manager? When Gizela, the aforementioned lab manager,...terminology of a new field. Annotate: When creating important documentation, including SOPs or project-related...Product Manager role at Addgene is “Words are important!”  If you try any of these approaches in your ...propensity for cross team communication, that’s an important soft skill to carry into the next phase of your...

...any other important features that differentiate this plasmid from its predecessor. Important features ...restriction sites used during cloning, and most importantly, the predicted full plasmid sequence. Other information...sequence verify the plasmids. This is the most important step in ensuring quality, as it will catch any...sometimes have to design new primers to sequence the important features. You can use primer design software such...

...gene incorporated into their genome. While transgenic mice have the new gene construct incorporated randomly...embryonic stem cells. Non-homologous recombination incorporates this DNA randomly into the mouse genome. In ...homologous recombination. A targeting vector is electroporated into embryonic stem cells and this will insert...vectors used to create knockout mice include a reporter gene, which replaces the targeted gene and provides...

...around the world, the spike protein has had many opportunities to evolve. Several variants are now circulating...different spike protein variants that can be incorporated into pseudoviruses. The lab used these pseudoviruses...Addgene to better understand which human genes are important for SARS-CoV-2 entry into human lung epithelial...). In their search, they found mucins have an important role in viral entry, which are inhibitors of SARS-CoV...research through sharing reagents has never been so important.     References and resources References ...

...experiments are important for visualizing ncRNA localization and function, but directly incorporating very long...transcribed but do not encode proteins. They play important roles in many cellular and regulatory processes...complex and not yet fully understood, so it is important to carefully select the target locations of knock-in...nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily block the transcription machinery of a target...

... sections as a primary research article, it’s important to understand that presenting your work in poster...certain posters resonate with you (or not). It is important to reflect on these impressions when thinking ...concise as possible, communicating only the most important results to your audience. With a poster, you have...particular styles recognized to work well. It is important that your readers can navigate your poster and...prominently, making it the star of your presentation. Portrait. This style displays a poster vertically rather...

...these tough issues is essential for building a rapport with these communities, which is in turn crucial...epigenetics. So, I sought out a number of different opportunities to broaden my training beyond the biological...policy-related topics, as time permits. Also seek out opportunities to communicate science to non-technical audiences...either giving or running these events will be important both in preparing you for future work in these...communicate your work to a non-technical audience is an important skill for every scientist. After all, scientific...

...expression levels, the terminator also plays an important role in RNA processing and contributes to variability...polyadenylation. The addition of the poly(A) tail is important for stability of the mRNA, protection from degradation...degradation, and is integral to the nuclear export and translation processes as well.   Figure 2... non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and/or use...the Mason–Pfizer monkey virus constitutive RNA transport element lead to enhanced HIV-1 Gag expression ...

... I can support that.” That was interesting. If people could be against GMOs as food but support GMOs that... up for real: bank account, business cards, incorporation, laboratory lease, etc. I still flip to that...result in a new utility, was there a business opportunity out there for us? After about a month of discussion...color, but the experience is invaluable. This opportunity has given us the chance to develop the technology...

...builds on existing popular web services, currently Crispor and Crispresso, that adhere to the same principles...decided to adapt the popular guide design tool Crispor by Max Haeussler. With a few extra lines of code... to analyze  CRISPR repair outcomes, which is important for quality control. The only ones we found that...analysis because of its ease-of-use and comprehensive reports. With CrispyCrunch, you get the all information...coding region in a UTR, CrispyCrunch flips the two important base pairs of the PAM (NGG -> NCC, CCN -> GGN)...

...detector using a series of mirrors and filters. Importantly, conventional flow cytometry uses one detector...broad, detectors will usually only pick up a small portion of that spectra to use for compensation.   ... flow cytometry for a given experiment, it is important to consider how many parameters you want to use...conventional and spectral flow cytometry, it is important to consider the expression levels of the markers...two highly overlapping cellular markers, it is important to pair them with two fluorophores that have distinct...