We narrowed to 886 results for: URE
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TypeBlog PostPublishedSept. 8, 2015, 2:30 p.m....phosphorylation, promoting transcription (see figure below). Subsequent exposure to red light rapidly deactivates ...limited studies and can fluctuate with changing culture conditions. The use of low intensity light solves...promoting transcription of a GFP reporter. Subsequent exposure to red light rapidly deactivates the system. Version...systems The large dynamic range of these systems ensures that they can be used with many types of proteins...Scouras A, Ellington AD, Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the...
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Using AAV for Neuronal Tracing
TypeBlog PostPublishedAug. 9, 2018, 1:04 p.m....understanding the brain’s function is knowing its architecture, in particular the connections between different... uptake of these dyes in vivo, microinjection, pressure injection or iontophoresis (applying small electric...of RABV. This improvement resulted in a clearer picture of neuronal connectivity in the mammalian brain...but overall with rather low efficiency. Not all measures of transport efficiency are consistent which may...observations like these may be a starting point for future developments in the field of AAV vectors. Conclusion...achieve transgene delivery by targeting peripheral structures (such as injection into muscle) as opposed to...(2008). Concepts in the pathogenesis of rabies. Future Virol 3(5), 481–490. PubMed PMID: 19578477. PubMed... -
Q&A with Karen Katz and Carl Paratore, the Newest Members of the Addgene Board of Directors
TypeBlog PostPublishedFeb. 19, 2019, 1:36 p.m....important considerations for the future of open science? What about the future of open science are you most...do you hope Addgene will accomplish in the near future? Katz: I’m excited that Addgene is continuing to...those “what could go wrong scenarios” and making sure we have adequate process and controls in place to... particular area or profession, you need to make sure you have learned and mastered the fundamentals. ...was a founding member of Mass Medical Angels, a venture mentor at MIT and served on the Boards of BioBuilder... -
Hot Plasmids - May 2022
TypeBlog PostPublishedMay 26, 2022, 1:15 p.m....powerful approach for proteomic mapping in both cell culture and whole organisms (see our 2019 overview). Until...doi.org/10.7554/eLife.64631 Goertsen D, et al. Nature Neuroscience. 2022. https://doi.org/10.1038/s41593... applies both positive and negative selective pressures for novel capsids. In a continuation of that work...Find M-CREATE plasmids here! Goertsen D, et al. Nature Neuroscience. 2022. https://doi.org/10.1038/s41593... factors, including limitations of number of flourescent proteins that can be inserted into a cell without...visual barcode plasmids here! Kaufman, et al. 2022. Nature Comm.https://doi.org/10.1038/s41467-022-30008-0... -
Antibodies 101: Monoclonal Antibodies
TypeBlog PostPublishedJune 29, 2021, 1:15 p.m....focus on antibodies that recognize proteins. Figure 1: Steps in the generation of hybridomas for monoclonal...fused to myeloma cells at a 1:1 ratio and then cultured in a special medium called HAT. This medium selects...antibodies are particularly useful when you want to ensure that the antibody binds to a specific region of... during your experiment, then you’d want to make sure that you choose an antibody that recognizes an epitope...polyclonal antibodies - which are a heterogenous mixture of antibodies that recognize many different epitopes...plasmid to create a recombinant antibody. Figure 2: Changes to the heavy chain or light chain genes... -
Fluorescent Proteins 101: Luciferases
TypeBlog PostPublishedJuly 22, 2025, 1:15 p.m....and alternatives to fluorescent proteins. Figure 1: Firefly, from Wikimedia Commons. The North...reaction in its abdomen to produce yellow-green light (Figure 2). Despite the title of this post, this reaction...photon; no chemical reaction takes place. Figure 2: Simplified chemical reaction catalyzed by luciferase...luciferase gene, and expression of luciferase is measured via the light produced. A similar approach can...luciferase or alternative substrates can permit measurements of luminescence from live cells. Luciferase ...with experiments that supply both substrates and measure luciferase output at different wavelengths. Addgene... -
Screening for Successful Genome Editing with Digital PCR
TypeBlog PostPublishedMarch 30, 2017, 2:30 p.m....negative (digital) signal at the end of the reaction (Figure 1). Partitions that did not contain any target ...precision. This means digital PCR provides an absolute measure of target abundance, does not require standard ...directly at the predicted double strand break site (Figure 2A). The single molecule resolution of digital...completely “blind” to this very important difference (Figure 3). If you are interested in harnessing the power...requires target DNA sequencing to confirm the exact nature of your mutations. A great description of the plethora...induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology 31, 822–826 (2013). PubMed PMID: ... -
Clear and Accessible Writing
TypeBlog PostPublishedFeb. 15, 2024, 2:15 p.m....densities are captured at the 17%, 25%, and 60% layers, and genome-containing virions are captured at the 40%...pun intended.) Steps to clear writing Let’s use Figure 1 as our example. Remember, our goal is to use ...same information as sighted readers. Figure 1: An image from AAV Purification by Iodixanol ...What are they hoping to get from it? The image in Figure 1 appears on our protocol page for AAV Purification...group to share with the rest of the workshop. For Figure 1, Addgene chose this alt text: "The left panel... -
Viral Vectors 101: Types of viruses
TypeBlog PostPublishedJune 6, 2023, 1:15 p.m....cell line cultures. Lentivirus Lentivirus is a type of retrovirus and shares many common features with gamma-retrovirus... gamma-retrovirus including genome architecture, host integration, and its association with disease. The...cell types. Altogether, AAV has many excellent features for human gene therapy and is currently being ...types of interest. This is a particularly nice feature as organ-specific injections, or the location of... Table 1: Types of viruses and their features Choosing a virus Now that we’ve reviewed the... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog PostPublishedFeb. 28, 2023, 2:15 p.m....provided by the manufacturers and enables customized experiments. Projects such as the OpenFlexure Microscope...modify the device for individual experiments. Manufacturers rarely provide the construction plans or CAD...accessible open-source hardware. Projects such as the OpenFlexure microscope or open-source modular optical toolbox...documentation, which facilitates the reproduction of the structure in a similar way to the biological protocol, and...et. al, "Robotic microscopy for everyone: the OpenFlexure microscope," Biomed. Opt. Express 11, 2447-2460... -
A Novice's Perspective on Neuroscience - SfN2017
TypeBlog PostPublishedDec. 15, 2017, 2:07 p.m....scientists and nonscientists alike). My only in-depth exposure to neuroscience prior to this conference was a.... This was not at all the case. The conference featured people who work in sociology, behavior, and artificial... session organized by Story Collider and which featured a talk about the different parts of the brain ...getting into neuroscience from the mol bio side you’re sure to have many opportunities to learn about and develop...website and on the Addgene blog to get some ideas for future experiments and to find tools that will make your...Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering Primer on Optogenetics Resources... -
RNA Extraction Without A Kit
TypeBlog PostPublishedMarch 24, 2020, 1:15 p.m....and work quickly to mitigate RNA degradation. Make sure to use DEPC-treated or RNAse free water. If using...water, autoclave the water to inactivate the DEPC. Ensure that plasticware or glassware used is RNase free...or more large, expensive ultracentrifuge. Figure 1: An outline of the different steps in RNA extraction...you need really clean RNA, it’s important to make sure that when extracting, to only take the aqueous layer... from) on your resuspended RNA sample will help ensure DNA contamination is not an issue. References...thiocyanate–phenol–chloroform extraction: twenty-something years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot... -
Transferable Skills Guide: Cross-team Communication
TypeBlog PostPublishedNov. 21, 2019, 1:53 p.m....the biggest project I’d seen in my already long tenure at Addgene. I would work with our research team...of what we discussed and decided. I needed to a) figure out the sources of miscommunication and b) find... steps towards solving this problem by 1) making sure we qualified these words when we discussed them ...Creating this guide was relatively easy. I took a picture of virtually every container we used, then for ...team identify possible points of confusion in the future. Improving cross team communication, be it in ...productive. Higher Productivity. Taking active measures in improving cross team communication has helped... -
Plasmid Tools for Microbiome Studies
TypeBlog PostPublishedJune 27, 2019, 12:25 p.m.... is limited and many microbes are difficult to culture outside of their natural environment. Thus, the...Pamela Silver’s lab developed a synthetic mark and recapture strategy for bacteria to count cell divisions....generations can be determined. The lab used DCDC to measure growth rate of E. coli for over 10 generations ...reveals growth dynamics in the gut microbiota." Nature communications 6 (2015): 10039. PubMed PMID: 26615910...engineering of the mammalian gut microbiome in situ." Nature methods 16.2 (2019): 167. PubMed PMID: 30643213... analyzing gut microbiota and host biomarkers." Nature communications 9.1 (2018): 3347. PubMed PMID: 30131493... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog PostPublishedJan. 31, 2019, 1:32 p.m....understood, a team of researchers lead by Takayuki Kogure (Kogure et al., 2011) believes that miRNA may decrease...metastasis." Nature 527.7578 (2015): 329. PubMed PMID: 26524530. PubMed Central PMCID: PMC4788391. Kogure, Takayuki...Microenvironments are cellular regions with specialized structures. Tumors are surrounded by microenvironments and...instead of repressing them, making it even harder to cure cancer. To understand how cancer cells influence... -
RANbodies: Reporter Nanobody Fusions
TypeBlog PostPublishedApril 10, 2018, 12:56 p.m....sequence, making them easy to harvest from cell culture media. Their His tags allow for further purification...for your RANBody While all three reporters stain cultured cells and tissue sections, some reporters are ...secondary antibody. Table 1: Summary of the Key Features of HRP, Spaghetti Monster, and Chicken IgY Domain...Tyramide signal amplification of HRP Staining cultured cells and tissue sections Amplifying weak signals...versions Labeled anti-HA or -Myc antibody Staining cultured cells and tissue sections When detected with a...(Y) Labeled Anti-Chicken IgY antibody Staining cultured cells and tissue sections Great for multicolor... -
Antibodies 101: Conjugation
TypeBlog PostPublishedMarch 26, 2024, 1:15 p.m....types. Be sure to check the spectral properties of the fluorophores you are considering to make sure that ... peptides, like an Fc-binding domain (Figure 1). Figure 1: Examples of sites on antibodies used...process quick and easy. Before getting started, be sure to check that your antibody’s buffer is compatible...2017) Simultaneous epitope and transcriptome measurement in single cells. Nat Methods 14:865–868. https... -
Hot Plasmids - October 2020
TypeBlog PostPublishedOct. 13, 2020, 1:15 p.m....tools to tease out the details underlying brain structure and function, with the advent of each tool expanding...excitatory and inhibitory synapses in the brain. Figure 1:Schematic diagram of different FingR constructs...protein products have distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different...more red than blue (Eastman et al., 2020). Figure 3: BaF3 cells were transduced with H2B-FT virus...which depend on IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation...proliferation and had a greater number of red cells as measured by flow cytometry. Cells grown in high IL-3 showed... -
How to Prepare for an Industry Interview
TypeBlog PostPublishedJuly 9, 2020, 1:15 p.m....process. Industry interviews, however, were more structured. I met with several individual and/or small groups...a theme too, which is something not inherently captured by the STAR method. I like to think of the theme...evaluations, professional development, and company culture. It’s also a good idea to prepare 1-2 unique questions... of your work, such as improving a process or procedure, solving a problem, or making a key decision. ...hours, which was a lot of extraversion for me. Make sure to get enough sleep the night before. Have your ... professional First impressions matter, so make sure yours counts. Dress professionally but still like... -
A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog PostPublishedMay 7, 2020, 1:15 p.m.... without DNA cleavage. Nature 551:464–471 . https://doi.org/10.1038/nature24644 Gaudelli NM, Lam DK, Rees... editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base...and dCas to the genomic DNA of interest Figure 1. When ABEs are in action they bind DNA so that...NG-Cas9 (PAM: NG) to create NG-ABE8.xm/d and the S. aureus Cas9 (PAM: NNGRRT) to create Sa-ABE8.x-m/d. With...increased activity and therapeutic application. Nature Biotechnology. https://doi.org/10.1038/s41587-020...