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We narrowed to 801 results for: Ran

Showing: 341 - 360 of 801 results

  1. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ...more detailed level, the Gal4/UAS system is a transcription activation system co-opted from yeast (1). It...workhorse to turn genes on or off. Gal4 is a transcriptional activator that binds to UAS enhancer sequences...sequences found in DNA. It then recruits transcription machinery to the site to induce gene expression. Thus...reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression as a means...15262411. 4. T. D. Southall, D. A. Elliott, A. H. Brand, The GAL4 System : A Versatile Toolkit for Gene ...Lai, T. Lee, Genetic mosaic with dual binary transcriptional systems in Drosophila., Nat. Neurosci. 9, 703...The Q system: a repressible binary system for transgene expression, lineage tracing, and mosaic analysis...

  2. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    Published
    Sept. 8, 2015, 2:30 p.m.
    ...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...types of proteins, including Cas9 and dCas9-transcription factors, for which low expression levels are...

  3. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    Published
    Dec. 18, 2014, 4:06 p.m.
    ...Wondering how you can develop more transferable skills? See Developing Transferable Skills During Science Training...of the University of British Columbia. When transitioning from an academic science career path to a non-academic...advantage over other candidates. Write about your transferable skills Some of the skills you developed might...research. For example, did you help put together a grant application? Are you familiar with the publishing...have presented in some public lectures? These transferable skills could be meaningful for your potential...Originally on track for a research career, she transitioned into a communications role instead after she...

  4. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    Published
    June 24, 2014, 3:59 p.m.
    ...codon, which is then transfected into the desired cells. A hybrid mRNA transcript containing luciferase...luciferase, usually serves as an indicator of transcription within cells, where detection of the reporter...Higher Lower Background Lower Higher Post-translational Modification Not Required Required Photobleaching...regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these...as the most efficient means to disrupt the cell membrane and deliver the substrate; however, secreted luciferase...luciferase and the gene of interest permits translation of functional luciferase, until miRNA target and degrade...

  5. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    Published
    June 1, 2017, 2:30 p.m.
    ...brightness When a dish of mammalian cells is transiently transfected with a plasmid containing a fluorescent...fluorescent proteins, e.g. see Chudakov et al (2010), Cranfill et al (2016) or Thorn (2017). However, the theoretical...developed in our lab uses a reference protein that is translated from the same open-reading frame and separated...reference protein) under identical conditions, a ranking of the practical brightness can be made (Goedhart...repeating this with another fluorescent protein, a ranking of practical brightness can be made. This strategy...the comparison of the practical brightness of two orange fluorescent proteins, differing by only a single...

  6. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ...synaptic transmission. Cleavage of VAMP2 renders it inactive, thus inhibiting neurotransmitter release....Plasmids podcast Light-mediated block of synaptic transmission Article contributed by Klaus Wanisch  Listen...enable VAMP2 cleavage, and suppresses neuronal transmission within minutes. Constructs using the photodimerizer...utility of light-mediated suppression of synaptic transmission in brain slices from mouse, and showed that ...recognized as important regulators of post-transcriptional gene expression across the majority of cellular...an unregulated fashion and thus a marker for transfection. The other (mKate2) is regulated in the 3’ UTR...

  7. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    .... elegans. These dsRNAs target specific mRNA transcripts for degradation. Despite how easy these experiments...often completely ineffective or results in low penetrant phenotypes that are hard to study. To overcome...target gene between two inverted T7 promoters. Transcription from each promoter progresses past the specific...podcast segment A fundamental approach to studying membranes, organelles, and other subcellular compartments...enzymes function across a much broader temperature range than BioID, which only functions optimally at 37...and C. elegans.   Find the TurboID plasmids here! Branon et al. Nat Biotechnol. 2018 PubMed PMID: 30125270...This tetramer cleaves chlorophenol red-b-D-galactopyranoside on the assay paper turning it purple, allowing...

  8. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...the filament from (a-b), showing the appearance and disappearance of 649-mDia1 or 549-CP (upper, middle...takes is a bit of chemistry magic to produce a wide range of useful molecules linked to that substrate. If... with SiR-CA (magenta) and outer mitochondrial membrane labeled with SNAP-OMP25 with Atto590-BG (green...separately with different dyes. b) mDia1 and CP appearance at the filament tip from (a). c) Intensity traces... probes are not very good at crossing the cell membrane. So, you should investigate your options and weigh...are typically 230–250 aa. Smaller size isn’t a guarantee that a tag won’t disrupt the function of the target...https://doi.org/10.1038/nmeth749 Strebinger, D., Frangieh, C. J., Friedrich, M. J., Faure, G., Macrae, R...

  9. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ... has recently published techniques to follow translation in living single cells. However, before CRISPR-Cas9...the Max Planck Institute for Brain Research in Frankfurt am Main, Germany. She presented a novel technology...to be invited to learn from them at the MPI in Frankfurt am Main and perform similar experiments. Visualizing...Both antibodies have a short antibody specific DNA strand added to them. Only if the proteins of interest...are in close proximity to each other can the DNA strands take part in rolling circle DNA amplification. ...microscope she likes to travel, discover new restaurants, go to concerts and bake. you can contact her...neurons reveals a mechanism for regulating its translatability." Science 343.6169 (2014): 419-422. PubMed ...

  10. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...Cas9 fused to the reverse transcriptase (RT), which induces two double-strand breaks and deletes the intervening...They further develop a version of SEND for gene transfer with pseudotyped viral particles, expanding options...enzyme from the Liu lab to target opposite DNA strands and specify a large intervening deletion. Not only...deletions and introduce epitope tags and RNA transcription start sites. The lab further demonstrated that...uses paired pegRNAs targeting complementary DNA strands to program large genomic deletions with or without...PEDAR is capable of correcting larger genomic rearrangements as demonstrated in a mouse model of tyrosinemia...

  11. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...their mutations corrected, and the T-cells were transferred back into the infants. Almost all patients have...repertoire (1). In this example, it was possible to transduce the T-cells outside the body and thereby correct...techniques may hold the key. In the simplest form, translating gene therapies from conceptual design to clinical...that carries a small, approximately 4.7 kb single stranded genome. AAV belongs to the family Parvoviridae...AAV vectors are also capable of infecting a wide range of cell types including but not limited to muscle...disease-associated mutations. AAV vectors maintain persistent transgene expression over many years in postmitotic, long-lived...targeting and enhances endonuclease delivery for double-strand break repair." Gene therapy 17.9 (2010): 1175-1180...

  12. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ... efficient double stranded templates, rather than the more efficient single-stranded oligodeoxynucleotide...generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent proteins... them to localize to the synapse and minimize extraneous fluorescence from other regions of the cell. ...synaptic maturation pattern of hippocampal adult-born granule cells over time that were not previously possible... al., 2020). Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations...lentiviral CRISPR library that targets human glycosyltransferase genes and other proteins related to cellular...

  13. CRISPR 101 eBook...en español

    Type
    Blog Post
    Published
    Jan. 18, 2024, 2:15 p.m.
    ...receiving the translation, Addgene contracted with a professional scientific translator, L. Denise Roche...101: Una herramienta de edición Addgene tiene una gran audiencia internacional, incluido trabajando con...101 eBook is now available in Spanish!  This translation was gifted to Addgene by Daniel Mejías Sánchez...Roche, to edit the translation. We are pleased to announce that CRISPR 101: Una herramienta de edición ...  Many thanks to:  Daniel Mejías Sánchez, PhD, translator  Denise Roche, PhD, MD, editor, ldeniseroche@...

  14. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ... mechanism to accurately repair harmful double stranded breaks, in which nucleotide sequences are exchanged...selection marker HSV-tk is used to select against random recombinants.   Design your targeting construct...selecting for cells in which the construct has randomly integrated into the genome, a negative selection...step will kill off many of the cells that have randomly integrated all or large portions of the plasmid...negative selection marker selects against improper, random recombination into a different locus. Correctly...contain the negative selection marker, but cells with random recombination may incorporate the negative selection...and animal models, their efficiency is very low, ranging from undetectable to 0.1%. In contrast, new genome...

  15. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...this surface”….where the complex is docked on the membrane of the endoplasmic reticulum. And graduate students...reported cleavage site on the complementary DNA strand. And this is why we love working with researchers...”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting physical...protein must have in order to scan along the double-stranded DNA, recognize a PAM sequence, allow the complementary...pairing between the guide RNA and the target DNA strand, and then activate two different endonuclease active...structure captures the displaced non-target DNA strand and shows the HNH nuclease domain swung around ...juxtaposition to its cleavage site on the target DNA strand. In addition, this model also displays the several...

  16. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...over for other applications, like PCR, cloning, transfection, or long-term storage. But what about those ...inverted terminal repeats. We use NEB Stable to transform and prep high quality DNA that contains these ... in a protein expression strain, we suggest retransforming the plasmid into a strain meant for DNA purification...prepping. Be sure to stay within the recommended range of the filter or you might inadvertently lower the...concentration Low copy plasmids produce fewer transcripts of their antibiotic resistance genes and can ...for high copy plasmids in standard growth strains range from 12-16 hours, but cultures with lower copy plasmids...to Quantify Your DNA Learn to Do a Bacterial Transformation Learn How to Perform a Diagnostic Restriction...

  17. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...mammalian cells can perform human-like post-translational modifications that bacterial and most yeast ...be produced through a single or dual-plasmid transfection, depending on whether the heavy and light chain...they’ve been separated onto two. You’ll first transfect your cells with your plasmid(s), and then wait... from dropping the tube they are contained in (Randolph et. al., 2018). At Addgene, we ensure the efficacy... Bisson L, Gervais C, Grazzini E, Durocher Y. Transient Gene Expression in Suspension HEK293-EBNA1 Cells...A. Pathak, Christopher J. Roberts, Prasad S. Sarangapani, Protein aggregation, particle formation, characterization..., https://doi.org/10.1016/j.cocis.2014.10.002. Randolph TW, Schiltz E, Sederstrom D, Steinmann D, Mozziconacci...

  18. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...C-to-U and A-to-I transcriptome-wide off target effects observed in whole-transcriptome RNA-sequencing, ...would convert adenine to inosine, which the translational machinery treats like guanine. This RNA editor...amount of off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target...deaminase ADAR2DD to deaminate cytidine and used dRanCas13b (catalytically inactive Cas13) to target the ...the RESCUE system’s ability to edit endogenous transcripts finding up to 42% editing efficiency. They note...therapeutic applications, the authors tested various dRanCas13b truncations that would allow packaging the construct...

  19. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ... that may have warranted their own chapters, and certainly chapters that could warrant their own books..., and he wisely makes no such attempt. He is transparent about his own biases--many of which will be shared...non-scientists, Zimmer’s observations will likely range from mildly surprising to genuinely shocking. Mainstream...he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in the identification...within this section. The GFP story has its own Franklin in Doug Prasher, who originally isolated the GFP...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...

  20. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...species. Discovering ways to culture, propagate, transform, store, and share all of these new organisms continues...Phenotype thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific...evolution because electric organs arose in different branches of the evolutionary tree at least 6-7 times and...mushrooms) but there are over 1,000 species. No transformation techniques are available yet. Polistes (Amy... a system using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all...One can inject plasmid DNA in the eggs to get transgenic animals. They might be immortal. Manihot esculenta...ReMOT control system (Receptor-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod...
Showing: 341 - 360 of 801 results