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

    Type
    Blog Post
    ...CRISPR/Cas inspired genome editing was initially described by Cong et al. and Mali et al. in January 2013...Nine months later, more than 1500 publications describe work to improve the tool’s specificity, orthogonality...orthogonality, and multiplexibility in various species and describe an assortment of applications. Despite the ubiquity...
  2. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...plasmid tools that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab... aeruginosa, Pseudomonas putida, Pseudomonas fluorescens, and Pseudomonas syringae. These vectors contain...system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta Biochimica et Biophysica Sinica. https...
  3. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

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    Blog Post
    ...instability-prone lentiviral vector plasmid in Escherichia coli Stbl3. 3 Biotech 3:61–70. https://doi.org...lentiviral expression vectors using reduced-genomeEscherichia coli. BioTechniques 43:466–470. https://doi.org...cloning strategy for efficient genome editing in Escherichia coli. Microb Cell Fact 19. https://doi.org/10.1186...
  4. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

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    Blog Post
    ...our AAV preparations. We recently published a description of the VGS process and use cases in Human Gene...ligation and tagmentation preparation approaches described previously require a preliminary double stranding...signature sequences in the data. Unlike previously described thermodynamics-based methods of serotype confirmation...
  5. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

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    Blog Post
    ...immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques,...Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and Multiple Antibody Staining. J Histochem...Addgene blog Antibodies 101: Introduction to Immunofluorescence Antibodies 101: Affinity Reagents Antibodies...
  6. The Importance of High Titer for AAV Transductions

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    Blog Post
    ... light responsive than fluorescent proteins required to get a good fluorescent signal. It may not be necessary...are often necessary to visualize expression of fluorescent proteins, for example, as many viral particles...
  7. R Bodies: Membrane-Rupturing Microscopic Tools

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    Blog Post
    ...bodies decorated with a fluorescent protein remain functional. Attaching the fluorescent protein fusions allowed... body dynamics. Second, we showed that these fluorescent R bodies can rupture E. coli spheroplasts (cells...
  8. Synthetic Photobiology: Optogenetics for E. coli

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    Blog Post
    ...Refactoring and optimization of light-switchable Escherichia coli two-component systems. Schmidl SR, Sheth... at Addgene. Synthetic biology: engineering Escherichia coli to see light. Levskaya A, Chevalier AA, ...Multichromatic control of gene expression in Escherichia coli. Tabor JJ, Levskaya A, Voigt CA. J Mol Biol...
  9. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

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    Blog Post
    ...protein. OptoNBs were also tagged with an infrared fluorescent protein tag which let the team detect its co-...revealed a blue light-dependent shift in mCherry fluorescent to the surface of the bead, suggesting that OptoNBs...OptoNB-SOS activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These ...
  10. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ...biologically relevant modification, or add in fluorescent or chemically reactive side chains, all of which...through a seminal paper from Peter Schultz’s lab describing a technique that allowed for intentional incorporation...of Phosphoserine into Recombinant Proteins in Escherichia coli. 12(21). https://doi.org/10.21769/BioProtoc...
  11. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

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    Blog Post
    ...27478040. The lung cancer mutant collection is described in this paper. Systematic Functional Interrogation...the genetic pathway reference collections are described in this paper Systematic morphological profiling...  The genetic pathway reference collection is described in this publication We also wrote an extensive...
  12. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...cytometry controls we’ll discuss are single color, fluorescence minus one (FMO), and isotype. You should always...controls like FMO or isotype controls, which we’ll describe a bit later. Single Colors Single color controls...(C) with APC-Cy7.   FMOs FMO stands for fluorescence minus one. It is used as a negative control ...
  13. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    Blog Post
    ...Harvard University. In my previous two posts, I’ve described the fundamentals of how to work with Drosophila...Drosophila as an experimental model organism. I then described the Gal4/UAS system used by geneticists to study...manipulations. You can produce some very beautiful immunofluorescent images like in this paper (2). Metabolic assays...
  14. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...positive effect on cancer treatment (Ref 1). As described in the presentation, starvation or the use of ...can find useful tips on in vivo imaging in our Fluorescent Protein Guide. Throughout the conference aspirin... with T cell receptor engineering (Ref 5)  as described by Blue Flame Addgene depositor Professor Michel...
  15. Kiran Musunuru on the Newest TALEN Genome-Editing System

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    Blog Post
    ...Cell, Musunuru, Chad Cowan and their colleagues describe a much more efficient tool for doing that disease...people unfamiliar with this new TALEN kit, can you describe it for us? Musunuru: TALENs are a type of genome...including FokI endonuclease domain as well as fluorescent proteins used to sort the cells after the plasmids...
  16. Antibodies 101: Conjugation

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    Blog Post
    ...either through a chromogenic or chemiluminescent reaction. Chemiluminescence, which is light produced by ...detect your antibodies using techniques like fluorescence microscopy, flow cytometry, or spectrophotometry...
  17. Plasmids 101: Modular Cloning

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    Blog Post
    ...specifically refers to the organizational schema described above, and not any particular cloning technique...cellular engineering goal, several of which are described below. Multigene circuits assembled through MoClo...relationship between genotype and phenotype is best described with “it’s complicated.” Applying MoClo Interested...
  18. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...should appear fluorescent if they were infected. While there may be some low-level fluorescent protein or...virus. Alternatively, if the virus encodes a fluorescent protein, the packaging cells could be examined...
Showing: 261 - 280 of 705 results