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Showing: 301 - 320 of 520 results

  3. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...allow for two sets of microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the locus...protocol for MMEJ-mediated knock-in of a GFP-Puro cassette into a given locus, just upstream of a stop codon...microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks are necessary for...for knock-in: one on either side of the GFP-Puro cassette and one in between the 5’ and 3’ microhomologies...of PITCh. The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and 3' microhomology and PITCh-gRNA...creates two DSBs in the PITCh plasmid to release the cassette, and an insertion-locus-specific gRNA cleaves ...and 3’ microhomologies are added to the GFP-Puro cassette via PCR, and this construct is inserted into the...

  4. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ... brightness is compared to that of EGFP which is set as 1. Some proteins are very dim (e.g. TagRFP657,...

  5. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...have become a powerful tool to address a diverse set of biological inquiries. References Ogura, Teru...death. The team used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed...for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct repeats which can be ... selects for those cells that have removed the cassette through a second recombination event between the...

  7. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ...difficult to use in a multicolor/multichannel imaging set-up. In contrast, smFP tags can be recognized by distinct...

  8. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ... used successfully in preclinical and clinical settings for ocular conditions. However, AdV was abandoned...responsible for an autosomal recessive, severe, childhood-onset form of blindness name Leber’s Congenital Amaurosis... of new capsid variants, designing expression cassettes to regulate and optimize transgene levels, and...

  9. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators, which...packaging limits (i.e., how many sgRNA expression cassettes will fit). Cre-dependent expression of the SPH...

  10. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...the day of your chalk talk Bring your own fresh set of erasable markers. You’ll have the colors you want...research? Do you have innovative tools? A novel dataset? Unique expertise? A novel hypothesis? Part of ...

  11. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ... the use of a given timer in many experimental settings. Monomeric fluorescent protein timers Since the... simplifies cloning and makes your experimental setup less cumbersome. 3. Portability across multiple ...

  12. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...be useful for your studies.  Find the plasmids to set up your screening systems at Addgene! Schematic...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...

  13. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...nature, they are largely underutilized in research settings — mostly a consequence of Cascade's complexity...

  15. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...This modular approach combines an interchangeable set of fusogenic and targeting components to reprogram...Every few months, we highlight a subset of the new plasmids, antibodies, and viral preps in the repository...

  17. Bioinformatics at Addgene

    Type
    Blog Post
    Published
    July 14, 2022, 1:15 p.m.
    ...working on one process at a time, and testing a small set of data each time a new process was added, we progressively...easily shared with others without any complicated setup. We created, tested, and debugged each of our pipeline...

  18. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ...that do not express the protein. With this control set, you should see your target protein IP’d in the positive...

  19. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    Published
    July 25, 2024, 1:15 p.m.
    ...antigen or an antibody depending on how the assay is set up. Then they block the capture reagent to prevent...binding to a primary antibody.  While the precise setup of the method can vary, typically users first coat...

  20. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...facilitates the use of prime editing in pooled settings with hundreds to thousands of pegRNAs. Chromatin-based...primer binding site, and many more. Once we had our datasets with all the different features and editing efficiencies...PRIDICT2.0 (Mathis et al., 2024). The additional datasets used in the training of PRIDICT2.0 make this model...
Showing: 301 - 320 of 520 results