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  1. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...development and therapy. Nat Rev Drug Discov (2017) 16(2):89-100. DOI: 10.1038/nrd.2016.238 Additional resources...drugs. While we often think of these two fields as separate, they are deeply intertwined. In this blog we ...Target identification Not all diseases are fully characterized or have a simple pathology. In these cases, ...
  2. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    ...incorporation of biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools...complicated entities with a large range of characteristics (color, brightness, photostability, maturation...attach important references that serve to further characterize a protein of interest (and we very much appreciate...simplifies the process of building advanced multi-parametric search queries.  Looking for a bright, monomeric...emission peak around 600nm?  Simply input these parameters and you’ll find matching FP suggestions. If ...
  3. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA... a mouse model, and its success is not always guaranteed. To learn more about this process check out our...
  4. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...photoconvertible fluorescent protein. Nat Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication...optogenetics. Even though GFP has been so thoroughly characterized, it turns out this protein has a few more secrets...
  5. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed...but with far fewer off-target effects! This characteristic probably explains why pC016 - LwCas13a guide...
  6. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ... enrich the microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol...production phase. However, this has not been well characterized as it is not a common event. What are exosome...exo-AAVs rely on differential centrifugation steps to separate cell culture supernatant containing exo-AVVs from... when compared to the conventional AAVs (AAV preparations were always normalized to their genomic titers...29296848. PubMed Central PMCID: PMC5728288. Schiller, Lara Timantra, et al. "Enhanced Production of Exosome-Associated...2018): 293-305. PubMed PMID: 29767307.  Wassmer, Sarah J., et al. "Exosome-associated AAV2 vector mediates...
  7. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...improve the brightness of previous Nano-lanterns by 2-4 fold and permit five-color fluorescent imaging! ...expressed in cells, generates much less signal than comparable fluorescent proteins. This weak emission hasn...! These eNL constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine...Nagai laboratory has a done a wonderful job characterizing Nano-lanterns and eNL for many different applications...Nano-lanterns requires a linear unmixing algorithm to separate the various colors. In fluorescent imaging, you.... Filtering the emission wavelength partially separates the signals of various Nano-lanterns, but it does...
  8. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach....   Once an antibody is found that shows separation between the wild-type and the KO population (...optimization can be performed to both maximize the separation and reduce any background staining. This can ...for a flow antibody, the YCharOS (Antibody Characterization through Open Science) initiative has now started... started producing flow cytometry characterization data for selected antibodies, using the knockout approach...antibody can be assessed through its ability to separate the cell lines based on expression of the target...
  9. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ... tabular HTML formatted output. Surprisingly only 2 tools provide FASTA output. A few notable tools allow...Nature biotechnology(2014). PubMed PMID: 24880488. 2. Hsu, Patrick D., et al. "DNA targeting specificity...years ago I was a part of a group (Biology of Host-parasite Interactions, Institut Pasteur, Paris) that changed...editing the genome of Plasmodium falciparum (the parasite that causes malaria). I attribute their success... is, however, not a focus of this blog post. I separate sgRNA design tools into database and de novo solutions... With respect to de novo sgRNA design, I again separate it into two categories. On the one hand we have... al., 2013, other tools have incorporated new parameters such as the Doench-Root score (by Doench et al...
  10. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    ...peer-mentoring groups at Addgene have gone virtual!   2. Practice public speaking For almost any science career...points for being able to just look at a paragraph and do a character count in your head.  Here are some sites...
  11. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    ...PMID: 30083608. PubMed Central PMCID: PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent...school classrooms, like gel electrophoresis to separate DNA or transforming cells with green fluorescent...in functional genomics research Learn how the Arabidopsis research center brings plant science to life ...
  12. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK...called transfection. Transfection is somewhat comparable to bacterial transformation (the introduction... a problem depending on the duration or other parameters of your experiment. Mammalian cells double at...reduces the rate of plasmid dilution but does not guarantee 100% transfection efficiency. Stable transfection...
  13. Antibodies 101: Monoclonal Antibodies

    Type
    Blog Post
    ...create a recombinant antibody.     Figure 2: Changes to the heavy chain or light chain genes within...the antigen-binding sites of the antibody - the paratopes - may change and impact the specificity and avidity..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P...Versus Polyclonal Antibodies: Distinguishing Characteristics, Applications, and Information Resources. ILAR...
  14. Plasmids 101: Cre-lox

    Type
    Blog Post
    ..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells... it, allowing for two separate recombination events to be controlled in parallel. Have you used the Cre-lox...consisting of two 13-bp long palindromic repeats separated by an 8-bp long asymmetric core spacer sequence... which can be located either on the same or on separate pieces of DNA. Both 13bp repeat sequences on a...forming a dimer. The two loxP sites then align in a parallel orientation, allowing the four Cre proteins to...maintained).  Translocation: If the sites are on separate DNA molecules, a translocation event is generated...Cre into cells of interest. What if I need two separate recombination events? One potential limitation...
  15. Bringing Sustainable Practices to the Lab: Recycling

    Type
    Blog Post
    ...the world’s population, we account for an estimated 2% of global plastic generation. Less than 9% of plastics... References Geyer, Roland, Jenna R. Jambeck, and Kara Lavender Law. "Production, use, and fate of all ...
  16. Tips for Improving Your Next Manuscript

    Type
    Blog Post
    ...easily understood and consistent in design and layout. 2. Be concise in your writing  When writing a manuscript...worthless. But simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts...
Showing: 341 - 360 of 486 results