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We narrowed to 767 results for: SCR

Showing: 141 - 160 of 767 results
  1. RaPID Detection of RNA-protein Interactions

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    Blog Post
    ...and IREB2, which post-transcriptionally regulate iron metabolism pathway transcripts. By using RaPID, Ramanathan...signal (~250-fold enrichment) when compared to a scrambled RNA motif control). BioID2 had no detectable signal...experiments RaPID negative controls include both scrambled RNA motifs and cells transfected with BirA* protein...changes in RC2H1 protein expression and post-transcriptional regulation of SM1v1-containing RNAs could play...IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE binding proteins...
  2. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

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    Blog Post
    ...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery...specifically edit methylation sites.  Step aside, transcriptionally active dioxygenase  When I started to work...by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains...in the genome and directly interacting with transcription machinery.   Well, I’ve got some good news...Hydroxylation, Active DNA Demethylation, Gene Transcription, and Memory Formation. Neuron 79, 6 P1086-1093...
  3. Synthetic promoter AAVs for cell-type specific expression in retinal cells

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    Blog Post
    ...synthetic promoters. Legend: TSS: transcription start site. TFBS: transcription factor binding site. Image from..., the Roska lab built synthetic promoters from scratch and tested to see if they drove cell type specific...conserved sequences that are upstream of the transcription initiation sites of at least two genes with ...promoters by repeating mouse cell type specific transcription factor binding sites interspaced with random...designed using stretches of DNA that were transcriptionally active in retinal cell types in mice. ...
  4. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    Blog Post
    ...spreadsheet Data Field Description Plasmid Name Use a descriptive plasmid name and match the... entry process On the Addgene home page, if you scroll over “Deposit” in the mega menu at the top of the... in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options ...does this plasmid do?” in <200 characters. A descriptive purpose focuses on the practical application ... Primary Vector Type The vector type should describe how your plasmid is intended to be used. Choose...
  5. Common Injection Routes in Mice

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    Blog Post
    ...off-target pokes to nearby organs. Restraint: scruffing Location: peritoneal cavity Gauge: 25–27 Volume...suspension to create xenograft models. Restraint: back scruffing or mechanical restraint Location: back or hind...but has a pretty low learning curve! Restraint: scruffing Location: throat Gauge (bulb-tipped feeding tube...to intermediate (tail vein) Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL Intermediate ... < 0.02–0.05 mL Beginner Subcutaneous Back scruffing or mechanical restrainer Back or hind legs; between...Anesthesia Nose n/a < 0.05 mL Beginner Oral gavage Scruffing Throat 18–20 (bulb tipped) up to 10 mL/kg Intermediate...
  6. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    Blog Post
    ...typically a Cas9 H840A nickase fused to a reverse transcriptase) together with a prime editing guide RNA (pegRNA...containing a primer binding site and a reverse transcription template (RTT). The design of the pegRNA has... we performed high-throughput ‘self-targeting’ screens with oligo libraries where pegRNAs are coupled ...cell contexts. Figure 1: High-throughput screening of pegRNA efficiency and building prediction tools...sequence-based prediction models from self-targeting screens such as PRIDICT/PRIDICT2.0 provide valuable insights...your target. Figure 3: High-throughput screening of the effect of chromatin on prime editing efficiency...
  7. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    Blog Post
    ...their coat protein to screen for binding to the target protein. The protocol describes going through two ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...enriched pool of sybody candidates are then reverse transcribed into cDNA fragments that are efficiently subcloned...The enriched sybody pools from the phage display screens are subcloned (using FX cloning) into the expression...interest. The Seeger lab used the ELISA technique described in their protocol because it is less susceptible...
  8. Anatomy of a Plasmid Page at Addgene

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    Blog Post
    ...plasmid, how will it be sent to me? Description: This describes how the plasmid or other product (kit...useful to me? Plasmid purpose: This is a brief description of what the plasmid was designed and used for...vector’s plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and ...they’ll end up on the N or C terminus of the transcribed and translated insert. If it is an empty vector... sequencing. Growth in Bacteria: This section describes how to grow the bacterial strain in which the ...
  9. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...Unlike promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic...resistance gene. These reporters allow you to easily screen or select for cells that are expressing your gene...interest at high levels. Unlike vectors that express screenable or selectable markers from a unique promoter,...your gene as they are both derived from the same transcript. Of course multicistronic vectors do not have...efficient translation of multiple genes from a single transcript. The two strategies most widely incorporated ...incorporated into plasmids for research purposes are described below. IRES Elements Translation in eukaryotes usually...
  10. Deaminet 2026: Breakthroughs in Base Editing, Deaminase Biology, and Therapeutic Translation

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    Blog Post
    ...a trade-off: overextension past the reverse transcriptase (RT) template, resulting in unintended insertions... implicate specific residues in the reverse transcriptase polymerase domain as likely determinants of ...‑target workflows often fail to capture newly described editing outcomes or meet emerging clinical requirements... (Nicola Osgood, UC San Diego) enables pooled screening of hundreds of gRNA–base editor combinations in...alleles are caused by premature stop codons. By screening 18,000 pegRNAs across all 418 human tRNAs, they...and clinical relevance. AIMers Deepak Shivalila described Wave Life Science’s GalNAc-conjugated AIMer platform...endogenous ADAR in vivo. Editopes Nina Papavasiliou described using short guide RNAs to recruit endogenous ADAR1...
  11. Lentiviral Vector Uses and Overview

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    Blog Post
    ...for transcriptional activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A...Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...both for targeted modification and genome-wide screening.   Lentiviral vectors in the clinic Since lentiviral... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014...
  12. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    ...functions such as epigenetic regulation and transcription by using locus-specific chromatin immunoprecipitation...worked extensively with enChIP systems. Can you describe this technology and its advantages? Fujii: In ...biochemical analysis of genome functions such as transcription and epigenetic regulation. To elucidate molecular...enChIP system using CRISPR technology. Can you describe this technology and some of its potential applications... any other technologies that you would like to describe?  Fujii: I believe that "Biology is Method", i.e...Hodaka Fujii lab's Addgene page and click the "Subscribe to Alerts" link in the Addgene Alerts box. ...
  13. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

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    Blog Post
    ...biochemical properties. In 1995, Roger Y. Tsien described an S65T point mutation that increased the fluorescence...backbones for constructing fluorescent fusions. Transcription Reporter: Placing GFP under the control of a...promoter in a given cell type. This type of transcription reporting was among the earliest uses of GFP... gene of interest, or expressed from the same transcript as the gene of interest, but after an internal... GFP are available. Others: We’ve really just scratched the surface of the potential applications for ...to identify particular cell populations in drug screens, to visualize micrometastases in nude mice in cancer...
  14. Tips for Writing a Good Cover Letter

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    Blog Post
    ...experience and skills would help you do the job as described. Let’s imagine you’re applying to a job that ... your cover letter, you can expand on that by describing how you developed and used a ddPCR assay to answer...start by looking at the requirements in the job description. For each requirement, ask yourself if you have...exactly what you would want to talk about! You could describe your organizational system, your rationale for...company/organization]. I am a [adjective that describes you] scientist with over [number] years of experience...the university]” Include keywords from the job description to help the hiring manager quickly make the connection...
  15. Hot Plasmids - August 2020

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    Blog Post
    ...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine ...your studies.  Find the plasmids to set up your screening systems at Addgene! Schematic of the evolution...their BioRxiv post, the authors generated and described several improved genetically encoded fluorescent...Takayuki Iwaki, Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion...activating expression of TCF4 while monitoring transcript and protein levels. Plasmids for multiplexed ...
  16. Plasmids 101: Codon usage bias

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    Blog Post
    ...create secondary structures in RNA transcripts Effects on transcription (remember, it’s not all about translation...locations or fail to make it through the entire transcript resulting in the production of nonfunctional ...every amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...translation - e.g. codon choice may interrupt transcription factor binding sites) As you might imagine,...algorithms and DNA synthesis companies such as IDT and GenScript host online codon optimization tools. Keep in ...
  17. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ...expression method enables us to quickly screen and investigate transcription factor candidates and the regulation...technique that uses a virus and the plant's post-transcriptional machinery to silence genes of interest (Unver...use of introns, which will be removed from the transcripts in the plant but not from Agrobacterium. Test...transient leaf transformations are a great help in screening candidate genes and speeding up research. Certainly...Budak. "Virus-induced gene silencing, a post transcriptional gene silencing method." International journal...
  18. Antibiotic Resistance: An Old Solution but a New Problem

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    Blog Post
    ...antibiotic’, very often we immediately imagine a prescription for a bacterial infection. While this post will...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...development of resistance include (Figure 1): Over-prescription of antibiotics (for example, giving antibiotics...not finishing the whole course of antibiotics prescribed to them This may seem counter-intuitive considering...The take-home? Always take your medication as prescribed. How do we know which bacteria are resistant ...infection is still effectively treated. We can prescribe antibiotics for the shortest time period possible...
  19. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

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    ...They rely on a viral protein called reverse transcriptase to convert their RNA genomes into DNA to be ... larger protein: PR (protease), RT (reverse transcriptase), and IN (integrase). These viral packaging ...the retroviral life cycle for a more detailed description). Since the integrated portion does not contain... cycle initially proceeds as usual. Reverse transcription of the transgene occurs in the cytoplasm, then... McCray, 2010). However, IDLV genomes are transcriptionally active, leading to transient expression of...Lentiviral vectors tend to integrate into transcriptionally active regions of the genome (Schröder et ...
  20. Antibodies 101: Introduction to Gating in Flow Cytometry

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    Blog Post
    ...SSC-H, or FSC-W/SSC-W, respectively. H, A, and W describe the shape of a histogram, which graphically represents... of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W results from...photocurrent vs. time. The resulting histograms is described through maximum current (H), the time the cell...depends on cell type and laser voltage. The voltage describes an electric potential that can be applied to the...comparing two markers simultaneously, we commonly describe the resulting populations as double positive (...that there are three parameters — H, W, and A — describing the intensity and duration of the detected signal...
Showing: 141 - 160 of 767 results