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Showing: 301 - 330 of 792 results

  1. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...editing system is regulated not only temporally with doxycycline, but it can also be regulated spatially with...these base editors Spatially and temporally regulated AAV-based genome editing In 2016, Jonathan Ploski... mediated genome editing system that could be regulated by doxycycline. Now, they’ve developed another... that CRISPR efficiency can be controlled by manipulating complementarity between the gRNA and target....

  2. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 7, 2017, 1:58 p.m.
    ...caused this glowing phenomenon wasn’t until much later when, in the 1960s, Osamu Shimomura studied the ...Washington state. Using these samples, he was able to isolate two proteins from the jellyfish’s photoorgans; ... absorbs that blue light and glows green. In the late 1980s, another researcher, Douglas Prasher, got ...expressed GFP in exogenous organisms (E. coli, and later C. elegans) for the first time. After Chalfie’s ...fluorescent proteins like GFP really are. In this compilation we’ll take a closer look at how they’re used,...

  3. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ...react and learn. All of this requires genes to be regulated dynamically.” To understand how that works in ...other brain functions, Konermann needed tools to manipulate the expression of specific genes in the brain...and genetically target them to specific cell populations. They demonstrated use of the LITEs to control...transcription in as little as 30 minutes after light stimulation. Under those optogenetic controls, gene expression...chromophores flavin-adenine dinucleotide and methyltetrahydrofolate, which are universally abundant. Second...

  4. Stepping into a Virtual Internship: My Experience at Addgene

    Type
    Blog Post
    Published
    Aug. 13, 2020, 1:21 p.m.
    ...interests are very clear, I had no idea how to translate them into real world careers. I figured this internship...but this never happened. I actually started at a later date and worked entirely virtually. Stepping into...world, I was worried about how my ideas would be translated through Zoom calls and Google docs. I pondered...Canva, and I was able to create cute and on-brand templates for these posts. Some of recent Meet an ... departments of the company. I asked questions relating to their roles at Addgene. I had a great time ...

  5. Bringing Sustainable Practices to the Lab: Innovations

    Type
    Blog Post
    Published
    March 7, 2019, 2:39 p.m.
    ...plastic you’ve saved. Repurposing regulated medical waste Regulated medical waste (RMV), which we collect...environmental impact of those processes? Who even regulates them? You may be surprised to learn that it hasn...Environmental Protection Agency, which started making regulations such as preventing manufacturers from dumping...and reused materials such as glass pipettes or inoculation loops. While single-use is now a popular theme...

  6. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    Published
    Aug. 13, 2014, 3:49 p.m.
    ...application we were looking at, magnetic isolation of circulating tumor cells, we found that the binding...Oxford is dedicated to generating new tools to manipulate biology based on molecular features found in ...the SpyTag/SpyCatcher system, based on a protein isolated from Streptococcus pyogenes that locks itself ...the context of cancer? Howarth: The capture of circulating tumor cells (CTCs) is going to have a big impact...

  7. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...on antibiotic plates.  Importantly, because the bacteria from which plasmids are isolated grow quickly ...easily make large amounts of plasmid to manipulate and use in later work.     How do scientists use plasmids...plasmids? Generally, scientists use plasmids to manipulate gene expression in target cells. Characteristics...

  8. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...antibiotic resistance marker. After transformation, plate the cells on medium containing the corresponding...contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing agar. Only cells... are auxotrophic for specific amino acids. When plated on media lacking the essential amino acid, the ... a toxic metabolite in gram-negative bacteria. Plating on sucrose medium will select for cells that contain...gene is used for negative selection on 5% sucrose plates. Exchange of the sacB gene for the gene of interest...

  9. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    Published
    Nov. 25, 2013, 10:12 p.m.
    ...specific, programmable platform well-suited for various innovative biological and translative technologies. Addgene...only deactivate the Type I-F system and are not translatable to the other CRISPR subtypes. Still, the large...Mali et al. in January 2013 (7, 8). Nine months later, more than 1500 publications describe work to improve...enabling access to multiple plasmids used for various platform applications. For more information on CRISPR/Cas...

  10. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    Published
    Oct. 20, 2020, 1:15 p.m.
    ... of the time because of relationships. It’s much harder to build relationships when you are more desperate...your hand at a video presentation? Most video platforms make it easy to record yourself. You can watch...experiences to share. The conversation builds a relationship for the future and the 1:1 format is good for...you will make good progress on both building relationships and thinking about future career options. 5....

  11. Meet the New Addgene Board Members!

    Type
    Blog Post
    Published
    April 28, 2020, 1:14 p.m.
    ...with regard to compliance with health authority regulations,” Baradaran says.  Baradaran’s path to science...is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function of all 20,... a genetic screen to identify the factors that regulate cell division. And that really got me hooked on...lentiviral microRNA-based system for polymerase II-regulated RNA interference. In the first two months since...also interested to see Addgene expand the sharing platform to other reagents in biology. Luk H. Vandenberghe...

  12. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...establish a next-generation sequencing (NGS) QC platform that allows us to provide scientists with complete...launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral vector (...ligation and tagmentation require a double-stranded template.  Using viral genome sequencing to verify the ...double-stranded species that could be used as a template for downstream applications (Berns, 1972). Using...the Addgene blog Read all of our viral vectors related blog posts Read our blog post on AAV titers Browse...

  13. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    Published
    March 26, 2019, 1:40 p.m.
    ...pAAV.hSyn-FLEX.iGABASnFR.F102G) is noticeably dimmer than iGABASnFR, and accumulates in the endoplasmic reticulum, but exhibits larger...GlyR IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR IRES EGFP...EGFP Chimeric channels for neuronal manipulation Recombinases Plasmid Serotype Name 50363  AAV5 .... It is useful for activity imaging of large populations of densely labeled neurons because background...  It is useful for activity imaging of large populations of densely-labeled neurons because background...

  14. Interview: Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering

    Type
    Blog Post
    Published
    March 4, 2014, 4:49 p.m.
    ...activity of two populations of neurons at once. Addgene spoke to Boyden about this latest optogenetics ...compete against each other. Getting two-color stimulation has been a holy grail of the field. The problem...selective for different ions. Right now, we can modulate electricity. If you could selectively let in calcium...Independent Optical Excitation of Distinct Neural Populations. Nathan Klapoetke et al. Nature Methods. 2014...

  15. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...switched on and off reversibly. This ability to modulate fluorescence allows scientists to observe single... (T203H) (1). wtGFP normally contains a mixed population of neutral (protonated) and anionic (deprotonated...T203H mutation in PA-GFP generates a chromophore population that is more neutral, causing PA-GFP to display...with blue light (1). Similar to PA-GFP, scientists later developed red fluorescent PA-FPs by performing several...photons achievable with photoswitchable dyes. Correlative light and electron microscopy (CLEM) is an imaging...Ryoko, Hideaki Mizuno, and Atsushi Miyawaki. "Regulated fast nucleocytoplasmic shuttling observed by reversible...

  16. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...site-specific recombinases, regulatory elements, developmental regulators, and lots of viral components... as Pichia pastoris). If your goal is protein isolation, then the MoClo Pichia Kit will get you started...developed to study the basic biology of and gene regulation in the opportunistic pathogen Acinetobacter baumannii... Molecular Tools to Study Gene Expression and Regulation in Acinetobacter Baumannii.” ACS Synthetic Biology.... “A Unified Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer ...

  17. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...1, to create a “cumulative wealth distribution” curve, or Lorenz curve. By calculating the area under ...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...less of a concern, but Addgene still recommends plating the transformation mixture on solid media. The ...The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library product for...

  18. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...vector production, assembled AAV vector particles accumulate inside the producer cells and purification protocols...gradients. The current purification protocols for isolating exo-AAVs rely on differential centrifugation steps...Figure 2). Since the development of the exo-AAV isolation protocol by Maguire and colleagues, other researchers...et al., 2017), and exo-AAV8 expressing human coagulation factor IX in liver in a mouse model of Haemophilia...et al. 2018). The structure of exosome AAVs is related to the use of microvesicles as a vehicle for drug...

  19. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    Published
    June 29, 2021, 1:15 p.m.
    ...Antibodies, 1999). This creates an immortal cell population that will continue to produce the specific antibody...or, are injected with the whole protein. After isolating the B cells from an immunized animal, those B ...step. Then, a limiting dilution is performed to isolate individual hybridomas and ELISA is used to identify...the selected hybridoma is expanded as a clonal population (Holzlöhner et al., 2017). Why choose monoclonal... Addgene’s antibody plasmid collection Read the latest on Addgene's new antibody project ...

  20. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...gamma-retrovirus is Murine Leukemia virus (MLV). Naturally isolated gamma-retroviruses have been associated with cancer...lentivirus and gamma-retrovirus are so closely related, their required components are very similar. While...heterologous envelope or a post-transcriptional regulatory element can be used for either type of retrovirus...infect humans, many mammals, and even the avian population. Some adenoviruses are known to cause respiratory...induce a lower inflammatory response within hosts. Relative to the other viruses discussed in this article...

  21. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    Published
    April 27, 2023, 1:15 p.m.
    ...mouse lines, you can access and manipulate an even further defined population of neurons. For example, say... workhorse adeno associated virus (AAV) shows relatively low retrograde transport efficiency in most serotypes...expression, detecting neuronal signaling, or manipulating any other genetically controlled function. By...Fig 3B). These specific neurons could then be manipulated (e.g., activated or inhibited) by any genetic...

  22. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ...engineers regularly use the platform, why scientists are using the platform, and tips for getting the most...from Nature Jobs Blog While Twitter is a popular platform for scientists, it’s not the only tool in a scientist... Facebook. For each of the three social media platforms, there’s some statistics on what percent of scientists...given you tips for using various social media platforms, and 3) helped you fit social media into your ...

  23. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ... on the properties found in the literature or tabulated data that summarizes properties of fluorescent...) respectively. The theoretical brightness is calculated by multiplying EC by QY (EC*QY, sometimes normalized...several important experimental conditions that are related to the imaging strategy and the sample. Practical...developed in our lab uses a reference protein that is translated from the same open-reading frame and separated...the assay is depicted in figure 1. The strict correlation between the fluorescent protein of interest and...

  24. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...splitting up a sample into thousands of physically isolated partitions (such as droplets or wells on a chip...edit. Since exploitation of NHEJ to generate translational reading frame-altering indels for “functional...wild type target in each sample can be quickly calculated (details in [2]) and can be used to quantify ...few thousand cells from single wells of a 96-well plate without prior quantification. Comparatively, mismatch...AnotherLabRat.     References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited human iPS cells without...

  25. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ... promoter, gRNA and pol III terminator. These relatively large cassettes (considering the gRNA itself ... vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from these ...deletions of other regions, such as noncoding RNAs or regulatory elements, to examine their function. Bacteria... of polycistronic gene organization, and its translation to CRISPR/Cas9 shows great potential for multiplexed...gRNA Multiplexing Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR...

  26. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...aren’t, so repeated cycles will favor accumulation of the latter products. As noted above, a single cycle...repair, as there is no requirement for a repair template (sister chromatid or homologue) or extensive DNA... repair should take less than an hour, thus a population of cells constitutively expressing a targeted...NHEJ-dependent repair (“pop-in” insertion) provided a template containing compatible overhangs is available. ...

  27. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    Published
    Feb. 4, 2014, 3:50 p.m.
    ...watch my webinar, “Not Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One...be a bit random because you never know which relationships will take off, or who your new contact will ...current organization but also important to build relationships outside your organization. The mentor in your... doing it again next month?” Some mentoring relationships grow naturally out of friendships or work interactions...other ways they can help you grow. Mentoring relationships can last for an hour or for a year. Some click...

  28. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...non-selective macroautophagy of the cytosol leads to accumulation of Pho8Δ60 in the vacuole, where it is cleaved...structures based on morphology. Autophagosomes are relatively easy to identify: double-membraned structures... GFP allows for tracking the localization or accumulation of pre-autophagosomal structures, autophagosomes...comprised of two tandem fluorescent proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive...induce autophagy, red but not green fluorescence accumulates in the vacuole. See Figure 2 for an example of...

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

    Type
    Blog Post
    Published
    Jan. 11, 2017, 3:30 p.m.
    ...better salary increases that those in less densely populated areas. This assumption does not appear to hold...not getting a raise. Meanwhile, in less densely populated places like the University of Kansas and the University...say, salaries are showing surprisingly little correlation with cost of living at the location of the institution...important step forward in helping the postdoctoral population continue to pursue academic work, and to enhance...scientific enterprise by reducing the postdoctoral population which keeps growing despite a lack of available...

  30. Antibodies 101: Conjugation

    Type
    Blog Post
    Published
    March 26, 2024, 1:15 p.m.
    ...low abundance targets. Check out our post on Biotinylation to learn more about biotin and its use in molecular...groups from cysteines, or carbohydrates from glycosylation (Dennler, et al., 2015). However, there are ...Endogenous carbohydrates added to antibodies post-translationally. 4) Modifications made to antibodies, such ...2015) Antibody Conjugates: From Heterogeneous Populations to Defined Reagents. Antibodies 4:197–224. https...The Basics of Western BlottingPlasmids 101: Biotinylation Resources on Addgene.org Unconjugated antibodies...
Showing: 301 - 330 of 792 results