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We narrowed to 580 results for: vit;

Showing: 161 - 180 of 580 results

  1. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    ...especially when done in vivo. There are also sensitivity issues when attempting to characterize mutations... the need for a faster and higher-throughput in vitro approach. Using data from The Cancer Genome Atlas...them had not yet been functionally validated. In vitro assay allows first-time functional annotation of...collection to screen individual perturbations in vitro using Ba/F3 and MCF10A cellular models. Ba/F3 and...

  2. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... H+ specific response cytoplasmic retention insensitivity to other ions In BRET, A mutant Renilla luciferase...mutagenized and screened for speed and voltage sensitivity. This hierarchical screen led to the generation...QuasAr2 possess mutations for improved voltage sensitivity and speed, and an endoplasmic reticulum export...construct, named CheRiff, exhibits high light sensitivity and fast kinetics. By combining both the voltage...produced luciferase is truncated and lacks enzymatic activity. The Luc plasmid (intronless) is useful for counter-screening...vector that enables protein expression in any in vitro translation system. The gene set was carefully chosen...the study of promoters with either weak or high activities (Bert, et al. 2000). To design pXPG, the authors...

  3. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...been removed/mutated to neutralize the nuclease activity. For strains that still carry this gene, there... purification protocol to remove endonucleases activity. For these reasons choosing the correct solution...any divalent cations like Mg2+, preventing the activity of DNase. While this is very important to preserve...temperature or at 4°C. Storing DNA: temperature and longevity You should consider a couple of more things when...

  4. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...contributed by Aliyah Weinstein Sparse Predictive Activity through Recombinase Competition (SPARC) is a fully...788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed by Leah Schwiesow...propidium iodine (PI), allowing for calcineurin activity measurements in stressed or aging cell populations...al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial Cell, 2020. https...

  5. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ... a voltage sensor AAV Historically, electrical activity in the brain was measured with implanted electrodes...reporter allows a way to monitor neuronal electrical activity through imaging. Since it’s illuminated by red...fluorescence and resting fluorescence. It is useful for activity imaging of large populations of densely labeled...fluorescence and resting fluorescence.  It is useful for activity imaging of large populations of densely-labeled...

  6. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    ...this episode of Hot Plasmids! Light controls the activity of a microtubule-associated protein to regulate...quantification of base-editing and Cas9 cleavage activity Article contributed by Angela Kaczmarczyk  Listen...fluorescence is activated when APOBEC-Cas9n-UGI-mediated activity edits out a frame-shift mutation within the mCherry... utility in quantifying multiple gene editing activities. Click here to find the ACE reporter plasmid...

  7. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...motivated by a particular interest in monitoring the activity and biochemistry of living neurons in an effort... calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes...approach he and his colleagues took to optimize sensitivities can now be applied to dozens of other published...concentrations than calcium, requiring greater sensitivity. He and his colleagues came up with a solution...

  8. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...increase activity, enhance binding between the template and primer binding site, increase processivity, or ...prime editor sequence to improve expression and activity In addition to the numerous innovations that streamline...mutations in Cas9 previously shown to improve nuclease activity (Chen et al., 2021). In a system as complex as...R221K N394K Cas9 mutations Improve Cas9 nuclease activity ~ ~ ~ ~ ✓ ✓ ✓ Evolved reverse...

  9. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ... expertise, publications, and plan for future activities; including the interest of the greater scientific...one that you can pay for, but one that you are invited to join (selected due to expertise in field). Publicity...journal editors to request to be a reviewer. Proactivity is very important in this case.  You can still...it as documentary evidence for your case – the invitation to be a peer reviewer needs to be addressed to...least 3 full-time people engaged in researcher activities. The EB-1B requires that you provide two out ...

  10. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    ...study tested AAV infectivity after a range of pH exposures. They showed that AAV infectivity was not altered...alter the infectivity of the virus. However, there was a substantial reduction in infectivity after 3 weeks...

  11. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...polymerase, which has 3′→5′ exonuclease activity. T4’s polymerase activity can be thwarted if no dNTPs are available...to polymerize, allowing the slower exonuclease activity to proceed. By including only one type of dNTP... reading Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Li MZ, Elledge...

  12. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...WYL1 increases RspCas13d and EsCas13d cleavage activity The majority of Type VI-D loci contain accessory... by RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins...proteins may be broader regulators of Cas13d activity. This property makes WYL proteins an intriguing counterpart...anti-CRISPR proteins that negatively modulate the activity of Cas enzymes, some of which are also functional...

  13. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ... without causing high background due to cross-reactivity. When to use polyclonal antibodies One advantage...labs. Polyclonal antibodies also have a high sensitivity to the target protein due to their ability to...or fixation (IHC). Because of their increased sensitivity, polyclonal antibodies are particularly useful... epitopes, increasing the likelihood of cross-reactivity with other proteins. As with any experiment, ...

  14. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...2020. PubMed PMID: 31937940. Detection of neural activity using a red fluorescent genetically encoded voltage...detect and output the voltage changes in neural activity when paired with appropriate fluorescent optics...combines the Cre/lox system with optogenetics. Cre activity is controlled by red light by splitting Cre and...bringing each terminus of Cre together to restore Cre activity. As soon as red light or PCB is no longer present...

  15. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...induced and quantified by measuring the amount of radioactivity in the culture supernatant (this is indicative...degraded) and it is slow. Alkaline phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline...to generate an active enzyme. Pho8 phosphatase activity or the molecular weight shift of Pho8 from its...regardless of its localization in the cell. SEP’s pH sensitivity means Rosella will fluoresce green unless it’...

  16. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...identified seven genera in in vitro experiments, possibly because in vitro conditions did not optimally...antibiotic selection. The team tested this system in vitro and in vivo. They identified cells from 19 genera...showed that this method can be used for both in vitro and in vivo studies. Find distributed cell division...

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

    Type
    Blog Post
    ...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery ...how can a DNA-binding protein with no enzymatic activity cause demethylation?  Dividing cells depend on...Mattei AL et al. (2020) TETs compete with DNMT3 activity in pluripotent cells at thousands of methylated...conserved C-terminal domain of TET1 to regulate TET1 activity and function in development. eLife 7:e34870. https...

  18. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...Why study neural connectivity? One of the early lessons many of us learned in biology is that the body...address this, molecular tools to map neuronal connectivity are widely used in neuroscience. In this post...emerged as powerful tools for visualizing neural connectivity (Nassi et al. 2015). The recombinant viruses...of a G-deficient rabies virus recombinant: an in vitro and in vivo study. J Gen Virol. 2000 Sep;81(Pt 9...

  19. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...majority have been shown to exhibit more limited activity than SpCas9 (2). In many cases this is due both...alternative approach is to modulate Cas9’s nuclease activity by modifying the gRNA with which it is complexed...capability remains intact while its nucleolytic activity is eliminated (Fig. 1) (1, 3). The gRNA can be...means of exerting tight control over Cas9 nuclease activity while eliminating the reliance on orthogonal Cas9...

  20. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...no-G PAM library to enrich for Cas9 variants with activity on non-NGG PAMs, generating xCas9 3.0-3.13. To...xCas9 in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...PAM flexibility might also increase off-target activity. Surprisingly, off-target effects across 5 gRNAs... as improvements at NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct NGG PAM NGA...
Showing: 161 - 180 of 580 results