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We narrowed to 321 results for: POR C

Showing: 161 - 180 of 321 results
  1. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...mutations in the common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development...1172-1179. PubMed PMID: 28671695. 6. Münch, Robert C., et al. "Off-target-free gene delivery by affinity-purified...basic research and other ongoing clinical trials support the utility of AAV as a broadly applicable gene...
  2. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...Most FP fusions for FRET studies are at the N- or C-terminus of your target protein, but even these modifications...can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition...for observed FRET efficiencies between 80% and 10%. C Cy3NTA and Cy5NTA bind His tags similarly but with...nonspecifically soak up your fluorophore package. One important concept in FRET experimental design is that the...donor first where labeling specificity is most important. Donor labeling and the need for specificity Donor...protein molecules contain the donor and, more importantly there is no fluorescence background resulting...in the ribosome, resulting in site-specific incorporation of the attached fluorophore. However, since ...
  3. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...in mice. Scientific Reports 9: https://doi.org/10.1038/s41598-019-41088-2 Magnus C, Lee P, Bonaventura...10.1021/cn500325v Goutaudier, R., Coizet, V., Carcenac, C., & Carnicella, S. (2019). DREADDs: The Power of the...Targeted and specific delivery becomes especially important for studies in neuroscience where specific tissues...neuron need to be targeted. Targeting is also important in the development of therapies when expression...Picking Ligands Picking the right ligand is also important for building a chemogenetic system. Your experiment...cellular response you aim to manipulate. It is important to consider that the same receptor can elicit ...drug that only binds to one receptor, so it is important to fully characterize the compound, or compounds...
  4. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ... CRISPR plasmids deposited with Addgene by year. (C) Cumulative number of laboratories that have deposited...smoothed using locally estimated scatterplot smoothing. (C) Number of requests per year for pooled libraries,...plasmids/year from 2017 to 2023. This growth is supported by new labs joining the Addgene CRISPR community...year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized by ...plasmids deposited by country; square size is proportional to number of plasmids deposited by each country...plasmids requested by country; square size is proportional to the number of plasmids requested by each ...Barrangou for his authorial contributions to, and support for, this and previous Addgene CRISPR publications...
  5. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ....2307/1542146 Lazarus, J. E., Warr, A. R., Kuehl, C. J., Giorgio, R. T., Davis, B. M., & Waldor, M. K..... D., Jamal, S., Wistrand-Yuen, P., … Forster, A. C. (2018). Engineering a palette of eukaryotic chromoproteins...Characterization of the Pink and Blue Pigments of Pocilloporid and Acroporid Corals. The Biological Bulletin 189:288...chromoproteins and fluorescent proteins, it’s important to note that despite their distinction by name...interactions with cytosolic proteins. This step is important as non-specific binding to proteins can inhibit...improved expression in living cells. Scientific Reports, 9(1). https://doi.org/10.1038/s41598-019-48604...
  6. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...these methods a try.  References Højland Knudsen C, Ásgrímsdóttir ES, Rahimi K, Gill KP, Frandsen S, ... CRISPR reporters in the near future.  SRIRACCHA (stable, but reversible, integrated reporter for assaying...histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct...Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems provide ...Recently scientists have created CRISPR fluorescent reporter assays to quantify targeted Cas9 cleavage, and...and non-homologous end joining (NHEJ). In these reporters, editing typically results in a change in the ...this blog we will focus on two fluorescent CRISPR reporter methods, SRIRACCHA and GEMCherry2. We will cover...
  7. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...Maddalo D, Manchado E, Concepcion CP, Bonetti C, Vidigal JA, Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de...Scientific Reports paper introducing the delivery method in 2014, Manuel Goncalves lab report that AdV-...the likelihood that donor DNA will interact with sporadic double-stranded chromosomal DNA breaks “that always...
  8. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    Published
    July 14, 2017, 2:30 p.m.
    ...Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, T. J., Johnston, J. L., & Tan, S. (2017)....Generating DNA Molecular Weight Markers. Scientific Reports,7(1). doi:10.1038/s41598-017-02693-1. PubMed PMID...
  9. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...References and resources References Chadeuf G, Ciron C, Moullier P, Salvetti A (2005) Evidence for Encapsidation...Lindenbaum P, Martin-Fontaine M, Broucque F, Robin C, Hebben M, Merten O-W, Blouin V, François A, Redon...production process? These questions are especially important when producing multiple vectors in parallel because...
  10. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    Published
    March 6, 2018, 1:49 p.m.
    ...temperatures (up to 95 °C) and is active over a range of elevated temperatures (37–75 °C). T4 DNA ligase ...PMC4737175. 22. Liu, Pingfang, Artur Burdzy, and Lawrence C. Sowers. "DNA ligases ensure fidelity by interrogating...commercially available (see below). Conclusion It is important to note that thermostable, high fidelity, nick-selective...
  11. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ...Discovers America. New York./  Callahan, K., Peterson, C., et al. 2022. External collaboration results in student...Evolution and Development. 24(5):127-130. Kimball, C., Powers, K., Dustin, J., Poirier, V., Pellettieri... outcomes and the opportunity to make new discoveries. Provost notes the importance of acknowledging student...history. Interestingly, he also adds to the disproportionately large number of honorees to have trained ... and he wanted to provide the same kinds of opportunities for his students. But he also found that his...Bringing his research into the classroom. I had the opportunity to talk with Provost recently about his development...Undergraduate Research Experience (CURE). Having incorporated a discovery-based lab on planarian regeneration...
  12. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...conformation capture (3C) based methods, such as 5C or Hi-C (Han et al., 2018). Think of 3C methods as taking ...other 3D chromatin methods such as ChIA-PET or Hi-C, by increasing both the number of unique chromatin...but enhancers and chromatin looping play important supporting roles. Enhancers are cis regulatory DNA ...networks’ of 157 super enhancers.  3. Measuring temporal changes in chromatin interactions A third application...application for CAPTURE 2.0 is detection of temporal changes in chromatin conformation, like those that ...interactions. The chromatin story told by CAPTURE 2.0 is important, but it’s not the only plot line in gene regulation...Multiplexed capture of spatial configuration and temporal dynamics of locus-specific 3D chromatin by biotinylated...
  13. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ... epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest on the C- or N-terminus. ...Their porous surface provides a large area for binding, but antibodies conjugated within the pores are ...used for IP. These proteins bind only to the Fc portion of the immunoglobulin, leaving the antigen binding...or a magnet (magnetic beads), providing a solid support while unbound proteins are washed away.  While ...to be adjusted depending on the sample. It is important to remember that sample lysates will also contain...
  14. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...predictable kinetics over 14 hours of expression in C. elegans embryos. However, this initial timer was ...timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus embryos. Of course, timers...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably... your experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al. showed...fluorescent timers that change color from blue to red report on cellular trafficking." Nature chemical biology...Chronology of islet differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868...
  15. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume of virus ...by ddPCR begins with diluting the virus. It is important to note that the dynamic range of the ddPCR is...PCR plate. Maintaining a high droplet count is important for calculating the concentration of your sample...separation (Witte et al., 2016). This is particularly important if your amplicon is longer than 150 base pairs...
  16. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...overhangs Cas13a (C2c2) 2x HEPN crRNA RNA 3' A, U, or C (not required by all orthologs) Specific RNA cleavage...spacer sequence and consists of a single A, U, or C base pair. In bacteria, once Cas13a has recognized...of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in the pool...activity of Cas13a becomes activated and the RNA reporter will be cleaved resulting in activation of the...in the field and as always, we appreciate the opportunity to share these invaluable reagents with the scientific...
  17. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ...https://doi.org/10.1096/fj.01-0995fje. Tan, E., Chin, C. S. H., Lim, Z. F. S., & Ng, S. K. (2021). HEK293 ...doi.org/10.3389/fbioe.2021.796991. Urabe, M., Ding, C., & Kotin, R.M. (2002). Insect cells as a factory ...of production, saving about a week of work. An important note is that this is only possible because baculoviruses... will come down to which differences are most important to you — virus type, cost, time, or reagents and...
  18. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...construct. Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the... Golden Gate assemblies feature an inverse proportionality between the complexity of the assembly (number...The future of assembly DNA assembly methods are important tools for many areas of science, and researchers...
  19. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome interactions) DNase-seq DNA...commonly think of DNA as a linear polymer of A,T,G, and C nucleotides arranged in a double helix. People are...genome. But not all of those 13,000 regions are important for maintaining ESCs. So which of these 13,000...next-generation sequencing to measure the relative proportion of guide RNAs (gRNAs) across the population. ...
  20. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the affinity ... tags, a His tag can be fused to either the N- or C-terminus of a protein. Unlike other epitope tags –... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the... are available, but scientists would need to incorporate their recognition sites into their protein tag...
Showing: 161 - 180 of 321 results