Skip to main content
Addgene

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

Information

In Category

We narrowed to 577 results for: Vit

Showing: 181 - 200 of 577 results

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55...cells and one which diminishes the endonuclease activity of the enzyme. In vivo, only nascent NgAgo protein...can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA...does however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one ...

  6. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...find people who are. I wanted to do fun science activities with the public, so I reached out to people who...what career suits you best I began viewing my activities as types of experiments. I searched for new opportunities...made me happy. Grad school can provide plenty of negativity, so I volunteered at a place that made me happy...recharged. The fun change of pace helped break up the negativity and allowed me to focus on the positive. I went...

  7. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...28b-Cas9-His from Alex Schier’s Lab. gRNAs can be in vitro transcribed from ssDNA, which can be generated by... advantage has allowed scientists to perform in vitro and in vivo CRISPR studies in experimental systems...approach using RNPs has recently been described in vitro, with the hope of eventually using this method for...enables efficient protein-based genome editing in vitro and in vivo." Nature biotechnology 33.1 (2015): ...

  8. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...knockdown, it still displays substantial off-target activity. By mutating residues in ADAR2 that interact with...the RNA substrate resulted in 15% C-to-U editing activity. They then performed sixteen rounds of directed...ADAR2DD to further optimize cytidine deaminase activity. While RESCUE can perform C-to-U edits, the authors...East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and...

  9. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ..., easily purified protein with easily measured activity, MDH is particularly amenable to in-class labs...   It is important to note that research activities in a CURE must be authentic – students should...give up the control embedded in “cookbook” lab activities (follow the recipe, and you get a predetermined...using a 6XHis tag, and compared their specific activities by measuring NADH production.   CUREs are...

  10. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...identified in Doench et al., 2014 to predict sgRNA activity. In this study scientists examined approximately...sequence features and adjustments that improved sgRNA activity and thus library performance. The Root and Doench...This library was designed to improve on-target activity while reducing off-target effects in human genomes...G., et al. "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9...

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

    Type
    Blog Post
    ...Specific RNA cleavage (followed by non-specific RNAse activity in bacteria) Cas13a targeting with a single...the pool of nucleotides, the nonspecific RNAse activity of Cas13a becomes activated and the RNA reporter...LwaCas13a does not display nonspecific RNA cleavage activity in eukaryotic cells. They arrived at this finding...East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and...

  12. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...been transformed. But caution: check for GUS-like activity of your tissue. Run a control with Agrobacterium...gene since plants can have endogenous GUS-like activity (Hu et al., 1990). We commonly use pSB161 - pL2...therefore good temporal reporters to measure promoter activity, for example. Useful positive controls could include...115. Hu, Ching-yeh, et al. "Intrinsic GUS-like activities in seed plants." Plant cell reports 9.1 (1990...

  13. Hot Plasmids - August 2020

    Type
    Blog Post
    ...extracellular surface) which enables tracking of protease activity with a fluorescent protein--no affinity purification...tested different target sequences for improved selectivity and found that the best motif was from the Lck...commonly used for in vivo biotinylation via BirA and AviTAG. pMAK contains the signal peptide of MBP, followed...followed by the desired nanobody (NB) sequence, an AviTAG, and 6xHis tag. The MBP signal peptide allows the...

  14. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...a little more complicated since basal neuronal activity in your brain region of interest could be low ...21716290. Pubmed Central PMCID: PMC3775282. 2. McDevitt, Ross A., et al. "Serotonergic versus nonserotonergic...PMC3150193. 8.Han, Xue, et al. "A high-light sensitivity optical neural silencer: development and application...M., et al. "Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans." Nature...

  15. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...virus type 1 integrase mutants retain in vitro integrase activity yet fail to integrate viral DNA efficiently...investigating HIV integrase mutants in the early 1990s (Leavitt et al., 1996), and since then, numerous variants...PMID: 18688286 PubMed Central PMCID: PMC2496964 Leavitt, A. D., Robles, G., Alesandro, N., & Varmus, H....

  16. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ... of any of these phosphorylation events on the activity of a given protein? Classical methods for studying...non-phosphorylated recombinant protein and perform in vitro kinase reactions. Often, however, researchers lack...or have difficulties with kinase specificity in vitro. The establishment of a system in which purified...pSer/phosphomimetic dual mutant) elevated kinase activity over 70-fold over dual phosphomimetic mutation...

  17. Plasmids for Stem Cell Research

    Type
    Collection
    ... reprogramming to cardiomyocytes using calcium activity as a functional measure of success. Mol Cell Cardiol...

  18. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...infect non-dividing cells is not restricted to in vitro cell culture as lentivirus-derived vectors are capable...promoter of the 5’LTR has been deleted to reduce its activity. A CMV or an EF1α promoter has been inserted in...have to be aware that there is often  background activity and it requires continuous administration of Dox...GFP encoded virus, and the reverse transcriptase activity assay uses qPCR to quantify viral RNA. All of ...

  19. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ... Cas9 activity in my stable A549 cell pools. Using this virus, it’s expected that Cas9 activity, guided...Cas9 (see tip 2). Pro-Tips 1. To quantify Cas9 activity using this assay, you really need to use FACS ...described in Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9...

  20. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...Griesbeck, Heim, & Tsien, 1999), monitoring kinase activity (Ni, Titov, & Zhang, 2006), quantifying antibody...structural changes due to this bulky insertion are inevitable. Most FP fusions for FRET studies are at the ...commercially available as Lumio Green and Lumio Red from Invitrogen (now ThermoFisher Scientific). These reagents...strategies. FRET experimental design lends itself to creativity and innovation, valuable commodities in any NIH...
Showing: 181 - 200 of 577 results