...with various spacer lengths from 30-84 nucleotides, and designated this system REPAIRv1. Using next-generation...introduced through non-homologous end joining (NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component...that interact with duplex RNA, they hoped to destabilize ADAR-RNA binding to decrease off-targets. Of ...

...are no exception to this effect -- meticulously designed experiments in labs around the world were put ...requests correlate with paper publication The dataset includes 11,426 plasmid requests over 6 years for 35 papers...into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential decay process we observed, while...model for each region using stochastic gradient descent in Pytorch, which gave us the maximum likelihood...

...they wrote the job description. You check off all the boxes - you have the desired degree, you have mastered... gaps in scientists' transferrable skills. In describing the limitations of the study, the authors noted...over email. Examine your current and past job descriptions. Scan through all the things for which you are...

...that’s suitable for PCR amplification using pre-designed primers. By using this library to infect an animal...isolate DNA from the infected animal and use the pre-designed primers to determine which version(s) of the capsid... (figure 4f) of the tTA inducer virus, but the desired sparsity of labeling is achieved with lose doses...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nature biotechnology...

... on single papers could be more important.  Any slides, presentations, recordings, and other materials...those who were unable to attend. We capture the desktop and audio recordings of the meeting using teleconferencing... ensure that the journal club is useful for the widest audience, you should get feedback from those who...served. I hope that with some of the information described above you feel empowered to start your own journal...

...those seen when using a plasmid backbone that was designed specifically for transient expression (find some...expression of viral constructs is a useful tool as it provides a means to quickly check that a construct is functional...3rd-generation transfer plasmids have a chimeric 5'LTR that includes a CMV or RSV promoter as well as a portion of ...those three genes plus tat. The envelope plasmid provides a heterologous envelope for pseudotyping and is...

... meliloti, Rhodobacter sphaeroides, R. capsulatus, Rhodopseudomonas spheroides, Salmonella spp., Serratia...Brevibacterium methylicum, Caulobacter crescentus, Desulfovibrio vulgaris, Erwinia carotovora, E. chrysanthemi...meliloti, R. leguminosarum by. viciae, Rhodobacter sphaeroides, Salmonella typhimurium, Vibrio cholerae, Xanthomonas...

...The hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. ...two pieces: a single protein, computationally described as a fusion of four Cas7 proteins and a putative...RNA editing methods — streamlining experimental design and its potential for clinical applications. Specifically...high-fidelity rate. Both Cas7-11 and PASTE open up multitudes of possibilities at the bench and potentially...

...pseudotyping allows these viruses to infect neurons. Besides altering the host tropism of your virus, there ...neurons. However, VSV-G’s broad tropism can be undesired for in vivo studies since there’s the potential...VSV-G is also toxic at high concentrations as described above and is inactivated by the complement system...proteins that can be used for specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al...

.... After months, or years, of grueling effort to design, fund, and conduct a research project, the vast...provide more than enough time for researchers to describe their science, and for viewers to decide if reading...is of interest. In his research, Scott Spicer concludes that “… the emergence of the video abstract genre...websites). WeShareScience is a social platform that provides links to the videos, keeping the copyright and...

...client might need help with the intricate task of designing a strategic vision for a portfolio of candidate...content-rich, easy-to-understand set of PowerPoint slides. What are some of the more challenging parts of...enjoy the challenges of healthcare consulting for decades, others go on to work with commercial teams at ...nature of the job and the ways to prepare for it. Besides those conversations, reading online healthcare ...

...whichever guides are closest to the transcription start site (TSS). We recommend targeting the guides to a ... you should also be ready to test three or four guides directed towards that gene as there can be a large...transcription. That being said, we generally pick guides by putting the promoter of the gene into a gRNA...

...the time of manuscript submission. The rationale includes increasing the ability of reviewers (and, once...issue by requiring ever more condensed methods descriptions, legends and limiting numbers of figures such...could be mitigated by on-line services such as SlideShare, Figshare and Dataverse. Just as Addgene helps... raise the alpha isoform of GSK-3 to the same (deserved) stature as the beta isoform. Follow him on Twitter...

...same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication) are unique because by...addition to the XFP in the construct, dBrainbow includes small epitopes fused to each XFP. Secondary antibodies...introduced using a plasmid construct called Zebrabow-GateDest containing an RFP, CFP, and YFP downstream of ...cell type. The Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt...

...cofactor binding proteins as described above. The smallest tags are peptides that are engineered to bind...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains...

...pathological developments (Vieira et al., 2019) Besides their ability to regenerate, the axolotl has several...scientific history, highly characterized genome, and widespread utilization, Arabidopsis has made significant...Arabidopsis genome project marked the beginning of widespread Arabidopsis genetics research development. Arabidopsis...allows for easy cloning of constructs specifically designed for use in Arabidopsis. There are also important...

...Applying for a Faculty Position: the View from Both Sides," are summarized below. Check Out Erik Snapp's ...You want people to focus on you and your ideas. Slides will be     distracting. What is the big question...feedback. Many of these documents are too vaguely described in other sources of guidance or the document examples...single malt whiskies, but I remember the days of modest postdoc salaries all too well. If there's a group...

...blot: we meet again. So useful, yet so finicky to design and optimize. Today we’ll cover the normalization...normalization can amplify other errors in experimental design and gel quantification. Figure 3: In a perfect...final blot. Quantifying the entire lane of protein provides a loading control that can be used for normalization...Antibodies 101: The Basics of Western Blotting Technical Design of a Western Blot Troubleshooting and Optimizing...

...expression to specific tissues and cell types. If desired, spatiotemporal control of transgene expression...systemic AAVs to find commonly used vector cargo designs and administration doses. Note: If capsid immunogenicity... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected..., B.E., Gradinaru V. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nature Biotechnology... prioritization for cell type-specific enhancer design. eLife,11, e69571. https://doi.org/10.7554/eLife...

...above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc fused...Reporters. The LumiFluor reporter constructs were designed to be packaged into a retrovirus for infection...BRET probe for imaging. These authors set out to design a better fluorescent protein that could be co-imaged...acceptor, the more powerful the FRET signal. In well-designed FRET-based biosensors, the intensity of the FRET...

...every item I tested was constantly tracked with barcodes and spreadsheets. It was great to be in the lab...industry positions. There were, however, some downsides to working in a large commercial company. In my...aspects of the company and gave me the freedom to design new projects and grow in the role. I understand... job different every day. I love the freedom to design and develop my own projects, but working remotely...

...end-joining, but will also efficiently ligate many undesirable structures, including substrates containing gaps...fidelity") better than T4 DNA Ligase (21, 25, 26). Despite this higher fidelity, Taq DNA Ligase can still ...possible NN pairs in the template. The probes were designed such that each possible pairing resulted in a ... high fidelity ligase, along with careful probe design and reaction condition optimization, will be critical...

...uncertainty of it all. Though I had spent the first few decades of my life following a very straightforward conveyor... harassment in academia or a large company who decides how they are going pursue regulatory approval for...their Local Engagement Playbook for step-by-step guides on how to engage in your local community.  Read...comprehensive resource developed by Dr. Kelly Singel which provides an overview of all available science policy fellowships...

...to be present and recognizable by your antibody despite the manipulation. Additionally, not all sample ...large deletion that removes the entire exon that codes for the epitope recognized by your antibody. You...specific, then you expect that the bulk of the peptides identified to be from the target protein. This...tuned for ready-to-use antibodies from Addgene Designing a recombinant validation experiment? Find plasmids...

...response element (RRE), an internal promoter, and the desired gene (transfer vector plasmid), 2) the HIV-1 genome...process is to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging cell lines to avoid recombination...

...update, we have added new sections to share and describe some incredible new CRISPR technologies developed...much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using...transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements...

...eBooks, the Antibodies 101 eBook is designed to be a helpful desktop reference. Each chapter brings together...an Antibody” quite useful. If you’re looking to design your first flow cytometry experiment, “Chapter ...

...When performed correctly, fluorescence microscopy provides a high contrast image in which a bright signal...specificity to the viewer. Finally, modern microscope designs may further exploit the unique characteristics ...one object in the final image. Over the past two decades a number of techniques have been developed that...resolution and decreased phototoxicity. Lightsheet designs that allow for multi-view imaging can image thick...

...been infected with a microbe, to trigger immune cascades that clear these infections. Anything that generates...immune response and initiates different immune cascades. In the research setting, antibodies of different...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ...

...occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween...for Halloween and how seriously we take costume design. In case you need more convincing, I’ve got just...to great customer support by dressing up as our Zendesk email system.  Our 2013 winners dressed up as ...

...single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation...assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission...also reveal novel pathways. The FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction...

...and the Addgene Help Center. A big part of this redesign was pulling all of our science options into a ...Addgene’s website easier to navigate, thanks to design choices informed by Web Content Accessibility Guidelines...word for it — check out all the new features and designs addgene.org! And when you’re finished, feel free...

...2022, from the launch of a new material to the redesign of a community resource. Now that we’re in the...antibodies. Everything in this collection, which includes both neuroscience and general-use antibodies, ...website saw 2.2 million visitors from 231 countries. Guides and protocols were our most popular content, while...

...Feng Zhang’s lab at the Broad Institute and was designed and cloned by Neville Sanjana. This fantastic ...your gRNA of interest to mammalian cells. It was designed to have increased lentiviral titer as compared...sampling of the research it has already been used for includes work probing mTOR signaling in cancer cells, deciphering...

...one-day conference aimed at the general public that provides an accessible introduction to various scientific...science cafes, and Reddit AMAs. Their website also provides a variety of short- and long-form written content... information about SITN: DayCon event website (includes archived content) Science in the News website ...

...part of the story. This year, we made significant strides in making science more accessible and reliable....supporting the tools in our collection, such as blogs describing viral vector serotypes, or hundreds of Data Hub...detailing antibody experimental results. We also redesigned our website to make it easier to find what you...

...of familiar flowers." This new x-ray technique provides scientist with a new method to explore the inner...of delicate plants and other organisms without destroying the original, which could provide a new tool ... collected by Darwin on his voyage. Previous destructive techniques were incompatible with the need to...

...process When I first saw the comprehensive job description for this co-op, I was immediately interested ...had stood out to me because it gave me a full description of the tasks I would be responsible for, in comparison...it relates to other teams at the company. This provides new (and old) employees with information on how...

...spatial biology enthusiasts. This dataset currently includes 1049 community-validated reagents from 20 contributors... And the dataset is rich. Experimental data includes reagent type, target species, target name, host... inspired by the open-software community when designing the IBEX Knowledge-Base, seeking to create an ...

...happening inside cells for at least a couple of decades. Green is the classic color, but fluorescent proteins... greater than any fluorescent protein earlier described. “Until our proteins, there were lots of fluorescent...illumination with far-red light. As the researchers described in Nature Communications last year, “The capability...