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We narrowed to 954 results for: APP

Showing: 261 - 280 of 954 results

  1. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...the co-op application process, my advisor encouraged me to keep my values in mind as I applied. Thinking...My name is Lauren Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst...s and leave. I was really devastated when that happened. I didn't know if I should even stay in science...in your career path and still find comfort and happiness there. Read all of the Meet an Addgenies features...connections with my coworkers that may not have happened if we hadn’t had these conversations. Now, I am...

  2. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ...and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...resistance genes.  It’s that simple! Good luck and happy editing!    Thank you to our guest blogger!Daniel...Addgene blog CRISPR101: Editing the Epigenome  Mapping the 4D Nucleome with CRISPR/Cas9 Resources at ...Xu X, Tao Y, Gao X et al. (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discov 2,...

  3. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ... Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells. To do...type specific expression in the retina.  Four approaches for designing synthetic promoters  To build a...reproducibly labeled retinal cells can be found here.  Applications for synthetic promoter AAVs Besides restricting...synthetic promoters have a few other potential applications: Using promoter combinations for targeted expression... promoters expressed in. The lab tested this application using one promoter to drive expression of Cre-mCherry... rates of labeling are ideal of gene therapy applications, promoters that produce sparse labeling would...

  4. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... to increase AAV transduction efficiency by approximately 15-fold (Chen et al., 2006). 3. Transduction... at 4 °C). After ultracentrifugation, virus may appear as a white pellet. The pellet can then be rinsed... This antibiotic selection can be performed approximately 72 hours after transfection and should give ...give rise to a surviving population of approximately 20–50% of the packaging cells after 1–3 days of selection... could be examined with a microscope and should appear fluorescent if they were infected. While there ...transfection, higher-level expression after approximately 72 hours is likely the result of successful ...

  5. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...to be less expensive than an ELISA kit. In this approach, viral RNA is first converted to cDNA and then...several different lentiviruses in parallel, this approach should be considered. Similar to the p24 assay... Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried...have integrated into a target cell’s genome. This approach can discriminate multiple integration events, ...underestimating the viral titer. The main drawback of this approach is that it can be quite labor intensive. Target...virus may want to choose more straightforward approaches such as the FACS-based or colony-forming unit...

  6. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair...phenomenon to the presence of terminal proteins capping the ends of linear double-stranded AdV genomes....double-stranded chromosomal DNA breaks “that always happen naturally.” These adenoviral CRISPR/Cas9 genome...and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral vectors Since... Adv vector delivery is often used, the CRISPR approach based on HDR has the advantage of being a potentially...Strategies for Tissue-Specific Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp ...

  7. Of Myc and Men

    Type
    Blog Post
    ...The strategy they landed on was to recombinantly append their proteins of interest with unrelated peptides...generate a reliable antibody for protein tagging approaches” was not part of the goal when Evan’s team generated...plays in human health and disease. But no science happens in a vacuum!  Munro and Pelham learned of these... flexible - it could now be used in way more applications.   Fig. 1 Making a Protein Tag (Abridged...their study, some of the newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So...proteins expressed from cloned genes: deletion mapping functional domains of Drosophila hsp 70. Embo J...

  8. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...your resume pulled to the top of the pile when you apply for such positions. Of course, if your goal is to...transition from an academic setting—especially if job applications require teaching experience or high numbers ...web site. Each position is for a specific, pre-approved project. Other companies, such as Novartis, have...s program. Once you’ve found a few positions to apply for, compare and contrast industry postdocs at those...not. The point is that each company has a unique approach, and some might suit your needs better than others...background in cell and developmental biology and is applying this towards researching neurodegenerative diseases...

  9. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...10 modules often rely on two-step hierarchical approaches using different Type IIS restriction enzyme specificities...transformation plate can be manipulated to result in appropriate levels of colony plating densities. Ligase fidelity...accuracy of assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any...researchers continue to test the limits of DNA assembly approaches with increasingly complex experimental conditions...Kucera is a Principal Development Scientist of Applications & Product Development focusing on DNA cloning...Optimization of Golden Gate assembly through application of ligation sequence-dependent fidelity and bias...

  10. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...and Mar. 1, 2020. Figure 2: A simple approximation of the effect of COVID-19 on data smoothed with...a 7-day rolling mean. While this global approximation already shows us a rough "COVID-19 effect" on...maximum likelihood estimates of each parameter. We approximated the data likelihood with multivariate Gauss-...the supposed end of lockdown), request activity appears to return to normal. The model fit these two discrete...is a Ph.D. candidate at MIT. His research uses applied machine learning and statistical methods for fundamental...fundamental scientific discovery and high-impact applications.       Alvin Hsu is a graduate student at Harvard...

  11. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...involves partitioning a PCR reaction mixture into approximately 20,000 droplets using water-oil emulsion technology...to create a water-in-oil emulsion containing approximately 20,000 droplets. Each droplet contains the material...droplets. In the image to the right, there is approximately 1 positive copy per microliter in the no template... use ddPCR for AAV titration, there are many applications for the technology.  Droplet digital PCR is ...being used in the detection of microbes. Some applications include measuring viable probiotics and detecting...detecting circulating pathogens.  With its many applications and ease of use, droplet digital PCR is becoming...

  12. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...a number of labs are using enChIP, and I'm very happy to see that many researchers all over the world ...describe this technology and some of its potential applications? Fujii: We generated a series of retroviral ...the peer-review system of journals and grant applications, for example. However, at the same time, it'...Throughout your career, you have held research appointments in the US (New York University School of Medicine...which only one was done in Japan, which is very disappointing. I hope I can help change the situation in the...and graduate students as they complete their appointments in your lab?  Fujii: These days it is very difficult...

  13. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...their experimental needs, and some of the common applications in the lab. Why Green Fluorescent Protein?...Re-engineering GFP to increase its range of color and application Almost as soon as its sequence was elucidated...the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising common...others) Neutral mutations    A multitude of applications Due to its size and ease of use, GFP and other...change. Typically, two fluorescent proteins with overlapping excitation/emission spectra are used; one fused... just scratched the surface of the potential applications for GFP. It has also been used to identify particular...

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

    Type
    Blog Post
    ...difficult to source a commercially available and appropriately conjugated antibody. While labs can conjugate...find,  it is often easier for labs to source appropriate antibodies for an indirect ELISA than for a direct... capture antibody must bind to distinct, non-overlapping epitopes so that they do not interfere with each...specific and can be difficult to develop if an appropriate conjugated antibody is not readily available....cross-reactivityChallenging to develop assay Competitive Appropriate for small moleculesReduced chance of cross-reactivity...choose the best ELISA for your needs. Good luck and happy binding! More resources on the Addgene blog Antibodies...

  15. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...This distinct approach means researchers can now select the best tool for their application from among many...incorporate A bases across from abasic sites. This approach resulted in a maximum A → T product purity of ...AYBEv3!). Although AYBE, AXBE, and ACBE do not approach the high product purity of CBEs and ABEs, they...). Several different teams succeeded with this approach around the same time (He et al., 2024; Tong et...on in the base editing field! From CRISPR-free approaches to mitochondrial base editing to base editors...

  16. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse...ligase cereblon (CRBN), including popular new approaches like PROTAC targeted protein degraders. But CRBN.... elegans embryo, where developmental processes happen within a few hours. In order to use the latest ...cellular processes in C. elegans: mScarlet3 for applications needing intense brightness (such as low-expression...simplicity and efficiency of this dual-plasmid approach makes it a valuable resource for advancing our...when the module unfolds under tension. When this happens, BiT and the LgBiT luciferase fragment (expressed...

  17. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...hour. Find the SHERLOCK plasmids at Addgene! Applications of the SHERLOCK detection system The Zhang lab...portability. This method can be used for an array of applications including clinical diagnostics (e.g. pathogen...While the lab points out that the protocol is not approved for clinical use at this point, they hope that...patient samples. Find the DETECTR plasmids here! Applications of DETECTR in diagnostics DETECTR allows for...an array of molecular and clinical diagnostic applications. The company Mammoth Biosciences has been started...SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing validation...

  18. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...cells retain the same color, allowing for the fate mapping of cell populations over time. The ability to track...variety of species, cell types, and research applications. Labeling neurons with Brainbow The Lichtman...neurons in the cerebellum of mice, the team appropriately named their technology Brainbow. Brainbow 1.0...level of these XFP is not strong enough for all applications. To resolve this, similar systems have been ...cell type when crossed with mice expressing the appropriate Cre allele. The Confetti mouse contains the Brainbow...PMC3713494. Ghigo, Clément, et al. "Multicolor fate mapping of Langerhans cell homeostasis." Journal of Experimental...

  19. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...Depending on the research question and downstream application, one or several of those techniques can be used...your flow analysis. However, certain downstream applications, like an in vivo adoptive transfer of those ...or free amines of proteins. Depending on the application, they can be membrane-permeant or membrane-impermeant...are positive or negative for the dye. The main applications of dye stains in flow cytometry are cell proliferation...with a high resolution for specific downstream applications, while maintaining a clean cell surface, dump...and bind DNA or free amines, depending on the application. Paul Heisig is a Research Associate in the lab...

  20. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...you with these, so keep reading! Good luck, and happy optimizing!  Recommended Reading Issa, S. S., Shaimardanova.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...A. (2021). Next Step in Gene Delivery: Modern Approaches and Further Perspectives of AAV Tropism Modification...AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways...Retrograde Serotype Viral Vectors 101: Viral Applications...
Showing: 261 - 280 of 954 results