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Showing: 451 - 480 of 1024 results

  1. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...efficiency constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for... binding of the plasma membrane. In previous publications, they described tools to control formation of.... Restricting the effect to a specific time or location can be critical when, say, the gene is essential...that trap the fusion protein away from its usual location, preventing it from carrying out its proper function...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...substantial PA accumulates in the plasma membranes, indicated by the PA probe colocalizing with SuperPLDx30....

  2. Tips for Getting a Faculty Position

    Type
    Blog Post
    ... do need multiple first/corresponding author publications in quality journals. It helps to come from the...a job. The currency of scientific success is publications and grants. 2. Write succinct and easy to understand...humbling. My conviction that I had a flair for communication was rapidly dispelled. She would ask me, "Is...succinct. Slide decks leads to the opposite and can suffocate an exciting idea in a swamp of unnecessary details...essential for human health. Failure to do so is implicated in several human diseases. Proper patterning ...disease treatment, etc., again in clear terms. "Implications for human disease" is vague. Instead, consider...interviews. For point of reference, I submitted my applications all by snail mail and only had phone interviews...

  3. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...the FOR website, the FOR Boston 2015 organizers indicate that the main question they wanted to focus on...oriented, scientifically minded, and able to communicate complex information - all traits you are likely...Publishing Panel brought in editors from a variety of publications as well as young PIs and all gave advice on ...blogging, write reviews, or write for a university publication, improving your general writing skills will help...academic publishing goals and make you a better communicator overall. Don’t Write in a Vacuum Your research...cognizant of this during my PhD. Submitting a publication doesn’t have to be like throwing it into a black...Science The Future of Research Website Additional Publications of Interest Can Be Found Here Resources at ...

  4. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...“Off-target effects” a vague catch-all term for unintended consequences in an experiment. For CRISPR ... example, if there are 10 putative off-target locations you can 1) determine if editing at those sites...sequencing. While this may be unnecessary for many applications, it is the only comprehensive method available...binding by Cas does not always correlate with catalytic activity at those sites, especially when DNA mismatches...For a candidate site-specific approach, mRNA quantification or western blot of putative off-targets would...you’ve tried everything listed here. For some applications, this is ok, but it still must be controlled... typically confound an experiment due to their location (e.g. in a gene in a similar pathway, a growth...

  5. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ... also been developed which don’t fit in either category, but those are a subject for a future post! ... GAL80 under different promoters and create sophisticated patterns of UAS-driven gene expression. To further... PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic... PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, Christopher J. Potter... PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, et al. “Improved and ... PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O, Potter CJ. The Q-System...expression of genes for metabolic engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res...

  6. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...sequences To design experiments for further plasmid verification Identifying plasmid features While our quality...promoters, selectable markers, tags, origins of replication, and fluorescent proteins that are already included...clue about where the gene of interest might be located. By easily using BLAST to check the translated ...reference sequences. Using Snapgene for additional verification experiments: Primer and restriction digest design...repetitive regions. These regions require additional verification experiments that can be partially designed using... plasmid verification...

  7. Walking beside Academia and giving it a high five: My transition from graduate work to a position at Addgene

    Type
    Blog Post
    ...talking to people about and communicating science. The path to science communication This may come as somewhat... I would ultimately want to go into science communication but thought that I would be best poised to share...PhD myself. To continue to foster my science communication skills, I took on TA positions, tutored fifth...largely my work developing myself as a science communicator that got me here! Thus far being an Outreach... one of my supervisors warned me, the swift gratification of publishing posts can be addicting. Even more...

  8. 10 Ways to Share Your Science!

    Type
    Blog Post
    ... Foundation Round Table. In any field, good communication skills help when it comes to finding a job. ...an argument for taking the time to hone your communication skills. So, where can a graduate student or ...opportunities than ever before for scientists to communicate about their work and their interest in science...your area or at your institution; the news or communications office is a good place to start. It’s not unusual...about science writing as a career, the Science Communication Program at the University of California, Santa... your research stories. Consider practicing communicating your research by guest blogging here, just leave...

  9. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ... perspective – a plasmid-containing cell must replicate the plasmid in addition to its own chromosomal...fully synthetic compounds. Antibiotics can be categorized based on whether they directly kill bacteria ...bacteriostatic); however, the distinction between the two categories may be a bit of a gray area as some bacteriostatic...Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin...Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes...and can be used in place of Ampicillin in most applications. Antibiotics vary in their sensitivitly to heat...

  10. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...fluorescent protein: identification of the molecular determinants of color and applications in fluorescence... be used as a fusion tag for a wide range of applications from traditional imaging to FRET and stochastic...492 nm, respectively), they gave this a teal classification rather than cyan and named the protein mTFP1...the fusion protein localized to their expected locations. An analysis of mWasabi gave similar results. ...fluorescent protein: structural characterization and applications in fluorescence imaging." Biochemical Journal...

  11. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    Blog Post
    ...scientific and technical training is that these educational paths prepare you for a vast selection of career...tech transfer, management consulting, science communication, policy and the diverse options in nonprofit...science is much like industry but the “product” is publications—the pressure to produce is the same You get ...and teach students Collaborations may be less “complicated” - for example, easy to initiate a collaboration...is different about non-academia? Scientific publications are a nice to have, not a need to have Must ...every 3-6 years—a short and exciting ride Complicated matrix structures are common Small integrated...

  12. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ... by the AAV. On a plasmid map, its sequence is located between the ITRs. It can be anything you like as...expression of your transgene to a specific species or location. Promoters can also drive different levels of ...sequences that can flank the transgene and are located between the ITRs. Transfer plasmids often have ... the expression of your transgene. The WPRE is located at the end of the AAV genome, just upstream of ...promotes RNA transcript longevity. The polyA is located at the end of the AAV genome, just upstream of ...

  13. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...infection, because this setup allows for experimental replicates for each of the gRNAs. The idea was to assign...cassette is removed, the effector is expressed. By truncating one of the recombination sites in the construct...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ... reporter (blue) and stained with PI (pink) to indicate cell death. Image from Diessl et al., 2020. ...https://dx.doi.org/10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining...protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison...

  14. Celebrate Open Data Day with Addgene’s AAV Data Hub!

    Type
    Blog Post
    ...these tools, which aren’t always easy to find in publications, so this is a much-needed community resource...resource of over 100 entries is driven by the dedicated community of data-contributing researchers who...is helpful to report the genetic background and catalog number or RRID if it exists. This is helpful because...exactly which strain you used so if they wish to replicate your experiment, they know exactly where to get...detection methods. For example, you can provide location, titer dose (Example: 1E13 GC/mL) and subject ... we’d like to give a shoutout to all of those dedicated scientists who have contributed to the AAV Data...

  15. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...degradation, blocking translation, or chromatin modifications of complementary gene sequences (Figure 1). ...silencing can be easily induced by exogenous application of long or short dsRNA. However, it is important...unmodified dsRNA elicits immunogenic reactions that complicate the effectiveness and interpretation of RNAi....have benefited from the ease of use in dsRNA applications. Figure 1: Schematic of an RNAi-inducing...between targets) or multiple target sequences are concatenated. hpRNA has been extensively used in plant research...selection reporter whereby survival itself is indicative of active gene silencing. We achieved this by...Internal Marker for Silencing Allows for Rapid Identification of Loss of Function Phenotypes. Plant Biology...

  16. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...genetic code of living organisms and its societal implications even before the discovery of genes themselves...explores the concepts of animal testing and modifications. The premise of the novel involves Dr. Moreau...2018) were highly successful in the box office, indicating the insatiable appetite of the moviegoers for...induce a double-strand break at the target genomic location. This is typically followed by non-homologous ...point mutations in the genome. They fused a catalytically impaired Cas9 protein to a DNA-editing enzyme...deaminase to introduce a C→T change at precise locations in the genome. This was soon followed by the adenine...us to always consider the ethics and broader implications of our research.  Want the Genome Editing Trading...

  17. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...off-target sites across the genome and pinpoint the location of mismatches based on the sequences of the genome... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...those relating to DNA structure (e.g. epigenetic modification, bulges) that may also impact off-target edits...cell-based methods are genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by...Genotyping Options Technique Detection Limit Application Advantages Disadvantages GUIDE-Seq 0.1% Cell-based... HTGTS Not reported Cell-based Identifies translocations Limited by chromatin configuration, produces...detection of KRAS mutations: diagnostic and clinical implications." The Journal of Molecular Diagnostics 12.4 ...

  18. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...the classical purification protocol involving iodixanol gradients. The current purification protocols for...particles accumulate inside the producer cells and purification protocols often describe a process that begins...lysis to release AAV particles with subsequent purification steps. It’s also possible for AAV vectors to... This is particularly useful for therapeutic applications in vivo where endogenous anti-AAV antibodies...Adrienn, et al. "Virus vector-mediated genetic modification of brain tumor stromal cells after intravenous...

  19. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ...fields and geographical locations. In addition to providing reagents and educational resources for scientists...formation. The quest for plant compounds to treat complications of diabetes Gloria Otito Izu, PhD Candidate...nephropathy, one of the major pathophysiological complications of diabetes mellitus. Recombinant human proteins... employing synthetic biology resources and microfabrication technologies to generate a self renewable ... in the river, take water samples at specific locations, measure fluorescent signals from the sensor, ...

  20. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    ...tropism, replication kinetics, and cell damage profiling of SARS-CoV-2 and SARS-CoV with implications for ... Another example of reagents that have broad applications but have proven useful to the study of SARS-...vectors VSV-G and psPAX2, which were used in a publication by Xiuyuan Ou, et al. that characterized the ... requested these plasmids from Addgene. This publication then resulted in the development of a new plasmid...production and distribution is possible due to the dedicated facilities and manpower that a repository provides...

  21. Trading Spaces - Visiting Scientist Edition

    Type
    Blog Post
    ...equipment sharing or a collaboration – it requires re-location of the scientist to the new institute where their...equipment! Preparing for your visit   Design and communicate a research plan Your time at the new research... and you are set up for success. If you are communicating with a professor or institute, ask if you will...out to your bench mentor (if you have one) to communicate your plan as well. Making logistical and funding...expect to generate data that will contribute to a publication? A grant? A patent? It is never too early to ...Resources Resources on the Addgene blog Fostering communication in remote collaborations Transferable Skills...

  22. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ...unique barcodes for each TF help streamline identification. While the Zhang lab controlled their multiplicity...single TF was delivered to each cell, future applications could deliver multiple TFs to each cell, allowing...processes. For help with library transformation and amplification, refer to the Zhang lab’s previously published...published protocol and its relevant modifications on the pooled library webpage. Disease models Good disease...generate, in part because of difficulty with replicating the specific dysregulation in the disease. For...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome Library...

  23. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    ...Collaboration and creativity are pivotal in writing publications/grants too. As described by several works (Wuchty...backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and acceptance...remotely. The authors stress the fact that “miscommunication, a lack of leadership, and inappropriate tools...approaches can lead to frustration, delay of publication, or even the termination of a project.” Take ...setting. Check out the blog post “Fostering Communication in Remote Collaborations” to get more ideas....Lewis ASL, Weyhenmeyer GA, O’Reilly CM, Lofton ME, Catalán N (2018) Ten simple rules for collaboratively writing...

  24. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ..., Nature Communications, 2019. PubMed PMID: 31451697. Genetically encoded voltage indicators for use ... many aspects of ApoB-LP biology, including quantification of abundance, size, and localization in vivo...Pubmed PMID: 23068583 Thierer et al, Nature Communications 2019. Pubmed PMID: 31366908 Adapting sigma54... by Brook Pyhtila Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when... excitation light intensities have made these indicators difficult to work with.  The Gottschalk lab has...

  25. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection...photon emission is measured resulting from the catalysis of a chemical reaction requiring luciferin, ATP...Background Lower Higher Post-translational Modification Not Required Required Photobleaching/Phototoxicity...blue light with a peak emission at 480nm upon catalyzing a chemical reaction with the substrate coelenterazine...compatible substrate must be provided for the enzyme to catalyze the light-producing reaction. Many luciferase ...

  26. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ...rounds of replication are initiated before the cell has divided. This means that genes located near the...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...

  27. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...detected with tyramide signal amplification. Tyramide signal amplification occurs when HRP converts a fluorescently...reporters (i.e. fluorescent dyes, HRP, etc. ) after purification. Antibodies are difficult for an individual ...culture media. Their His tags allow for further purification, if desired, but it's not required for their...some reporters are better suited for particular applications. Read on to learn more about each reporter or...Pros Cons Antigens HRP (P) Tyramide signal amplification of HRP Staining cultured cells and tissue sections...

  28. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...members of their pathways, but where the kinases are located, their signal strength, and how long their signals...and MEK1 is active or “on.” Dronpa’s color thus indicates MEK1’s current state (Green when off, dark when...domains were inserted at sites homologous to the locations they were attached to in MEK. The photoswitchable...kinases good for? There are a number of potential applications for photoswitchable kinases.  Below are a few...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...endogenous ERK, and its localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates...

  29. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    ...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... little correlation with cost of living at the location of the institution. Continuing efforts to gather...Director of Future of Research. He is interested in advocating for and with junior scientists, to help them ... Bring Increased Postdoc Wages Read about the Advocating for Science Symposium and Workshop Learn about...

  30. Antibodies 101: Conjugation

    Type
    Blog Post
    ...These properties allow for efficient signal amplification, making biotin conjugated antibodies good options...especially if you start to consider clinical applications). But there are a few other classes that we ...proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et al., 2007...also conjugate antibodies directly to beads for purification purposes.  Coupling up - antibody conjugation...added to antibodies post-translationally. 4) Modifications made to antibodies, such as inclusion of non-canonical...Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell...
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