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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 581 results for: Vit

Showing: 81 - 100 of 581 results

  1. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...detect PKA activity can be generalized to measure phosphatase activity, second messenger activity, and protein-protein...faster assembly and disassembly dynamics, higher sensitivity to light, and better distribution of clusters...biosensors for live-cell imaging of biochemical activity Article contributed by Angela Holmes Jin Zhang... imaging of the dynamic changes in biochemical activity within the cell. Their Fluorescence fLuctuation...Dronpa. The authors tracked protein kinase A (PKA) activity at the plasma membrane in living cells using a...different time points, the authors tracked protein activity while also capturing information about spatial...

  2. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ... make this STEM activity fun,” but rather “let’s see how we can make this fun activity relate to STEM....their expertise with the kids through some fun activities. If your school doesn’t have a Math & Science...parents from any background, it really helps if activities are run by parents who can answer some of the... a lot of kids, you’re going to want a lot of activities (try for a dozen!). The best approach may not...STEM.” Figure 1 shows the activities we had for my school’s Math & Science Night. There are plenty of places... places to get ideas for science activities. I did some crowdsourcing on Twitter and got lots of suggestions...Steam Powered Family  Biology and Chemistry activities from Quatr.us Foldscopes Steve Spangler Science...

  3. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...your in vivo work is to screen you gRNA in vitro first. In vitro screening is a simple and fast method for...selecting the guide RNA, validating its efficacy in vitro, and using it in mouse embryos to generate gene ...supplied by the Zhang lab. This simple website invites you to input the target region (approximately 500bp... many previously validated gRNAs at Addgene. In vitro screening for the best gRNA Figure 1: Validation...Validated gRNAs to Target Mouse Genes To begin the in vitro screening process, you should first develop primers...in vivo. Find Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9 gene editing in mice Now ...  in vitro screening...

  4. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...the approaches that can be used to determine selectivity of an antibody for flow cytometry experiments...well as a pragmatic approach to determining the selectivity of antibodies for other sample types and for ...For these reasons, it is essential to test the selectivity of your antibody. But testing is not one size...your experiment. An antibody that demonstrates selectivity for cell surface labeling may become less selective...knockdown to enable robust assessment of antibody selectivity. Additionally, it can be a real challenge to ...protein level at the timepoint studied Off-target activities of the RNAi Correctly interpreting your data...protein expression makes confirmation of antibody selectivity even more challenging. Correlation with RNA or...

  5. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...balance maximizing on-target activity while minimizing off-target activity, which sounds obvious but can...100nt window downstream of the TSS gives the most activity. Thus, a given gene will only have a dozen or ...the method of producing the guide (synthetic, in vitro transcription, or lentiviral delivery) can affect...unwise to make conclusions on the basis of the activity of a single gRNA, and thus diversity of gRNAs ...approaches. Avoiding off-target effects The off-target activity of gRNAs is important to consider. While the basic...mismatches that can nevertheless still lead to activity has been established, and can be used to identify... indicates that the consequences of off-target activity, at least for the experimental conditions used...

  6. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...CasPEDIA feature. Activity features: CasID CasPEDIA will display both the cis and trans activity of each entry...enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability ...strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the other hand, generates staggered...manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.). With this... an evolving database, and all researchers are invited to contribute to it! The CasPEDIA Consortium will...update entries as research progresses, but you are invited to reach out to suggest new entries or even new...

  7. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ... protein activity and have important roles during oxidative stress. To monitor this activity, Pedre Pérez...biosensors that detect new redox metabolites are vital for researchers to better understand the processes...assessed cFos expression. She will explore the connectivity patterns of these cells and the functional contribution... will allow us to better understand how neuron activity changes following opioid withdrawal, providing...excitatory and inhibitory neurons whose function and connectivity are poorly understood,” says Qiu.   Qiu developed...both optogenetics and chemogenetics to study the activity of parvalbumin (PV)-expressing neurons, neurons...

  8. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ...retained the on-target activity of wt Cas9 while decreasing off-target activity at known off-target loci...displays comparable on-target activity and similar or decreased off-target activity across the sites tested ...effects of CRISPR/Cas9 by altering the nuclease activity of the Cas9 nuclease itself. Design and testing...Created with BioRender.com. To decrease the electropositivity of the HNH/RuvC groove, Slaymaker et al. made...combination mutants for their on-target and off-target activity at multiple genomic locations in HEK cells. Further...

  9. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...and responsive voltage indicator with enhanced sensitivity for unitary synaptic events By Brian O’Neill ...ASAP5 selected for its improved kinetics and responsivity in screening ASAP3 variants. B) Electrical and... and optical recordings of spontaneous activity in human stem cell-derived neurons expressing ASAP5-Kv...regimes with fast activation kinetics and high responsivity near the resting membrane potential, ASAP5 is...and responsive voltage indicator with enhanced sensitivity for unitary synaptic events. Neuron, 112(22),...context-dependent responses and molecular patterns of selectivity. With 10 plasmids available as a kit, plus 21...Direct interrogation of context-dependent GPCR activity with a universal biosensor platform. Cell, 187...

  10. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...gRNAs in vivo, they should be screened for activity in vitro and then packaged into AAV for in vivo delivery...SPH mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators...developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell et al., 2019). This ...Bdnf is expressed early in response to neuronal activity (hence, “immediate-early”) and consequently regulates...immediate-early genes associated with neuronal activity, such as Fos, Egr3, and Arc. Thus, CRISPRa represents...preferable for individual gene activation in select in vitro and in vivo applications, the tools described above...

  11. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro, and they further...SpCas9 variants have improved activity on NAG PAMS and reduced activity on NGG PAMs relative to wild type...mimic aspects of organ morphology and physiology in vitro and have emerged as a powerful model for understanding... the enzymatic biosensor retains its catalytic activity.  As SLs are also known to trigger seed germination...

  12. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In vitro, ligases (notably T4 DNA ligase) are critical reagents...circularization of dsDNA (6). Less commonly utilized in vitro, Taq DNA ligase will ligate only nicks (5-8). Taq... applications, there cannot be any end-joining activity, and for others, there is a need for the exclusive...or qPCR-based methods to greatly increase the sensitivity of detection (14-20). LDR has also been extended...achieved through a combination of the intrinsic selectivity of the ligase active site and careful balance... that extends earlier studies through a high-sensitivity multiplexed format (21, 27). This methodology...Bruce Wallace. "Specificity of the nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989...

  13. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Provided 5% Inexpensive, simple Low-throughput, low sensitivity Sanger Sequencing Provided 20% (variable) Sequence-level...inappropriate for heterogenous populations, low sensitivity Targeted Amplicon Sequencing Provided 0.01%...including Digenome-seq (Kim et al. 2015) for in vitro analysis, IDLV for in vivo detection (Gabriel et...potentially toxic) Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with... background BLESS Not reported Cell-based (in vitro) Can be used on tissue from whole animal models,...throughout the genome. However, their decreased sensitivity means that the best option moving forward may...most investigators use single cell clones for in vitro CRISPR experiments. The likelihood that a single...

  14. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    ...outdoors. Taking a walk or a bike ride is a great activity for both exercise and clearing your mind. And ...And because the outdoors is so huge, these are activities that you can easily do while staying away from... your life. At Addgene, we’ve started virtual activities like team lunches, a virtual soup lunch, and ...kids' needs. Plan ahead. Think about and set up activities that kids can do more independently. For example...meals and snacks ahead of time.  Find learning activities for kids. For example, Skype a Scientist is now...research community. Set expectations for your productivity Set a routine and stick to it. Plan defined...

  15. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...off-target activity than BE3  85172  pBK-EQR-BE3  EQR-Cas9 (NGAG)  Lower off-target activity than BE3 ...donors in experiments simultaneously measuring the activities of CaMKII and RhoA during induction of structural...domain similarly allows for control of enzymatic activity. In addition, proteins can be “caged” by fusion...editing. For each base editor, they observed editing activity with a minimum efficiency of ~50% and confirmed...85177  pBK-YEE-BE3  SpCas9 (NGG)  Lower on-target activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3...pET42b-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3...pCMV-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity; mammalian expression vector Rees H, et al. Nature...

  16. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...current understanding of neuroanatomy and neural connectivity, providing valuable insights for neuronal communication...methods have been developed to study neuronal connectivity. These methods are less neurotoxic and better...). When using these viruses to mark neuronal connectivity a large majority of neuronal cells are labeled...improvement resulted in a clearer picture of neuronal connectivity in the mammalian brain (Wickersham et al., 2007...tracing with AAV vectors is the Allen Mouse Brain Connectivity Atlas, a project to map neuronal connections... study can be found on the Allen Mouse Brain Connectivity Atlas website. A similar approach was used by...connections that are functionally relevant, i.e. activity-dependent neuronal tracing. Apart from mapping...

  17. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    ...links the expression of gene III to base editor activity and editing speed and consists of the following...Figure 2: Phage-assisted evolution of base-editing activity. A plasmid-based genetic circuit in the bacteria...links the expression of gene III to base editor activity and editing speed. Created with BioRender.com....of TadA variants with much higher base editing activity compared to the original TadA-7.10 enzyme. After...showed increased off-target DNA and off-target RNA activity. To mitigate this drawback, the lab introduced...editor with improved Cas domain compatibility and activity. Nature Biotechnology. https://doi.org/10.1038...

  18. Tags and Other Markers

    Type
    Collection
    ...Recombinant Antibody Description Source Species Isotype Reactivity Recommended Applications PI Do you have suggestions...

  19. Neuroscience

    Type
    Collection
    ...Recombinant Antibody Description Source Species Isotype Reactivity Recommended Applications PI Do you have suggestions...

  20. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...recombinase activity were exposed to blue light. Cre-Vvd showed substantially improved activity as compared...permitting recombinase activity. Opto-Cre-Vvd is precise, demonstrating low sensitivity to ambient light, ...gene expression tools in and of themselves, their activity can be fine-tuned and precisely controlled through...authors demonstrate that CRISPR-Casɸ is active in vitro as well as in human and plant cells. Casɸ...
Showing: 81 - 100 of 581 results