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We narrowed to 1,005 results for: RING

Showing: 421 - 440 of 1005 results

  1. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...pyogenes (SpCas9), is truly an exceptional genome engineering tool. It is easy to use, functional in most species... cell stress, and RNA size distribution. When comparing LwaCas13a to shRNA, they found that LwaCas13a ...vivo RNA imaging. In short, the Cas13a molecule brings a much anticipated element of diversity to the ... the CRISPR field. We feel that the culture of sharing within the CRISPR community is an essential part...scientific community. Note: Cary Valley and Mary Gearing contributed to the update of this post. References...

  2. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...expand and exploit CRISPR, both of which we're featuring in today's blog. Cas7-11 Development and Discovery...alongside some creative (and significant!) protein engineering. They combined Prime editing with an integrase...potential applications. But they weren’t done engineering quite yet – surely, they thought, the integration...optimized the specific attachment sites via sequence engineering, enabling even more efficient integration activity...: Adding Precision and Flexibility to CRISPR Engineering CRISPR 101: RNA Editing With Cas13  CRISPR Cheat...

  3. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...studying chemical engineering. Mengtong "Tom" Duan is a PhD student studying bioengineering. They are both...smaller genetic footprint, and nonlinear acoustic scattering. The best candidate for this was the Anabaena...was sufficient to saturate expression. When engineering a cell line, cotransfect the mARG cassettes 2...acoustic reporter genes for real-time in vivo monitoring of tumors and tumor-homing bacteria. Nat Biotechnol...

  4. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...software. When you are happy with your selection, ordering the sgRNA is simple and easy. Several companies...but fret not! The key to saving time and energy during your in vivo work is to screen you gRNA in vitro...in mice to the day you have your F0 generation (barring life’s little accidents and stuff ups). This is...multiple genes by CRISPR/Cas-mediated genome engineering." Cell 153.4 (2013): 910-918. PubMed PMID: 23643243...Addgene Blog Get the Basics of Mouse Genome Engineering with CRISPR Validate Your Genome Edit Learn about...

  5. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...resulted from excision by uracil N-glycosylase (UNG) during base excision repair. Adding a second copy of the...off-target mutations than those derived from naturally occurring cytidine deaminases. Adenine base editors The ...compared to ABEmax. In an alternative approach, engineering and directed evolution of the TadA domain has...) This post was originally written by Mary Gearing in August 2016 and was updated by Jennifer Tsang...Y., Guan, Y., Siwko, S., … Li, D. (2023). Re-engineering the adenine deaminase TadA-8e for efficient and...applicability of base editing through protein engineering and protein delivery. Nature Communications, ...

  6. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ... require anaerobic conditions. In experiments requiring anaerobic conditions, proteins that bind endogenous...bilirubin (Kumagai, 2013). Next to these naturally occurring co-factor binding proteins, systems wherein a ...proteins can be accurately determined in vitro by measuring the fluorescence intensity of purified protein...usually dim relative to fluorescent probes, recent engineering efforts have increased the brightness of these..., Marieke, et al. "A FRET-based biosensor for measuring Gα13 activation in single cells." PloS one 13.3...

  7. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...proven useful in the engineering of riboswitches. Riboswitches are naturally occurring regions in the 5’ ...into the content of Addgene’s aptamer soup by exploring the development and applications of fluorescent...Warner et al., 2014); its length reduces artefacts during live cell imaging. Bunch of baby spinach, or BoBS...transcribed by RNA Polymerase III (5S, tRNA and U6) and measuring Corn fluorescence by flow cytometry, the Jaffrey...

  8. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...also ideal for signal amplification during these experiments. During detection, a biotinylated reporter... biotin. Thus something in the egg white was sequestering biotin from being absorbed. In 1940 Esmond E...can be added to either terminus of a DNA sequence during a Polymerase Chain Reaction (PCR) by using a biotinylated.... Biotin can also be enzymatically incorporated during PCR using a biotin labelled dUTP. There are multiple...

  9. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...then used to measure the frequency of each variant during the selection process. Each variant is assigned...of MnCl2. Disadvantages of this method include favoring of certain types of mutations, difficulty controlling...then both strands of the plasmid will be nicked during the first BbvCI digest and the whole plasmid will...direction, both strands of the plasmid will be nicked during the first BbvCI digest and the whole plasmid will...Douglas M., Jason J. Stephany, and Stanley Fields. “Measuring the Activity of Protein Variants on a Large Scale...

  10. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...unfamiliar, scFvs are often the building blocks for engineering proteins. So let’s get acquainted with scFvs....CD3 on T cells and a tumor-specific antigen, it brings T cells to a tumor site. ScFvs can also be fused...viral vectors like AAV, which could be useful for delivering scFvs that inhibit HIV.   References and Resources...Bruccoleri RE, Haber E, Crea R (1988) Protein engineering of antibody binding sites: recovery of specific...more about antibodies  Read about another reagent sharing platform for antibodies, the Developmental Studies...

  11. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...Proximity labelling relies on the labelling of neighboring proteins within a cell with biotin. Biotin, is...promiscuous biotin ligase that can efficiently label neighboring proteins The first biotin based proximity labelling...from the ligase and bind to exposed lysines on neighboring proteins. Thus to capture protein complexes, ...electron-rich amino acids such as tyrosine on neighboring proteins resulting in their biotinylation. The...The advantage of APEX is its speed, labeling neighboring proteins in minutes rather than hours like BioID...

  12. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ... nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. What you ...4740. You can find more info on pCri in Mary Gearing’s blog post on the system Easing the Protein Purification...Seq kit has been used in microbiology studies monitoring sequence dynamics under various selective pressures... can lead to surprisingly large fitness effects during adaptive evolution of antibiotic resistance." Proceedings...2016): 895-899. Choudhury, Rajarshi, et al. "Engineering RNA endonucleases with customized sequence specificities...

  13. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...post-translational modifications, identifies a staggering ~250,000 phosphorylation events in the proteomes... led to the eventual discovery of a naturally occurring phosphoserine-tRNA synthetase that serves as an...elongation factor that accepts Sep-charged tRNAs during translation. Many ‘flavors’ of Sep-OTS plasmids...pSer-containing GFP that can be used as a positive control during optimization experiments. This version of GFP will...dephosphorylate the pSer-incorporated protein, ideally bringing the phenotype assayed closer to that of the wild...

  14. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...How much do you need [for your lab start up]?” A hiring chair or dean will often ask a faculty candidate...community is the ultimate value proposition. By preparing a budget before hand, you’re saving the institute...turned down a faculty position because none of the offering institutes were able to provide the resources ...If the resources aren’t available, then you’re entering a win-lose situation that can ultimately set you... be afraid to walk away from a negotiation. If securing a faculty position is your goal, then a resource...

  15. Addgene is Stopping Use of X (formerly Twitter)

    Type
    Blog Post
    ..., and as of today, will stop posting, monitoring, and resharing content on X.  Addgene deeply values our..., TikTok, Facebook, and LinkedIn. We are also exploring other social media platforms with active scientific...in the scientific community. Addgene has been considering a gradual move off X as a response to major platform...

  16. Trends in CRISPR and SynBio Technologies

    Type
    Blog Post
    ...the recent Keystone Meeting "Precision Genome Engineering and Synthetic Biology". His reflections on the...scientific community. Data on CRISPR and synBio plasmid sharing In a recent review on CRISPR technology, for example...online user forums such as http://www.genome-engineering.org and http://www.egenome.org." At the Keystone...Keystone Meeting Eric presented a poster with data comparing Addgene global deposit and distribution trends...

  17. New FLIS Plasmid (April Fools 2022)

    Type
    Blog Post
    ...accosted by dozens of tiny black dots during our stay there. Yet capturing the flies is nearly impossible, ..., due to their small size and inconspicuous coloring.  FLIS plasmid Recently, the Phuels lab deposited...everywhere find and re-contain any specimens that escape during routine lab activities.  Here, we demonstrate the...

  18. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...impossible. With the genome engineering revolution came epigenome-engineering tools - zinc finger nucleases...enhancer regions (Mendenhall et al., 2013). By comparing gene activation when enhancers were active or ...Sauka-Spengler T (2017). Genome and epigenome engineering CRISPR toolkit for probing in vivo cis-regulatory... Mary Gearing...

  19. A Control for All Seasons

    Type
    Blog Post
    ...treated identically, used to assess naturally occurring random biological variability. If one were studying...species-on-species staining can be partially addressed with stringent staining protocols and the use of isotype-specific...standard protocol, but the primary antibody is omitted during the incubation step. Samples are incubated with...will briefly touch on.  Western blotting When comparing samples in different lanes in a western blot, ...controls address the spectral overlap that occurs during multiplexing. Compensation controls are single ...

  20. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...cell-type specific. If it is something you’re considering for your experiment, note that you cannot extrapolate...hardware types, construction, and testing used for delivering your light stimulus. This can make or break your...Photoswitching Proteins (Vol. 2173, pp. 21–51). Springer US. https://doi.org/10.1007/978-1-0716-0755-8_...2019). Cortical layer–specific critical dynamics triggering perception. Science, 365(6453), eaaw5202. https...
Showing: 421 - 440 of 1005 results