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We narrowed to 798 results for: Cre

Showing: 181 - 200 of 798 results

  1. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    Published
    June 4, 2019, 1:02 p.m.
    ...site-saturation mutagenesis, and medium-throughput screening, she increased the affinity of the sensor for serotonin...overwhelming response and were excited to see the many creative uses of fluorescent proteins. We’re happy to announce...thousands of protein variants simultaneously  With increasing sequencing power, scientists are able to quickly... protein-coding pathogenic human variants are decreased in protein abundance, possibly due to misfolding...neuromodulator, and evidence suggests that it is incredibly important for nearly every aspect of life.” Unger...system. “This bottom-up approach will allow me to create a fully controllable and reproducible set up,” ...explains. Developing an artificial leaf replica means creating a “leaf” with similar permeability of water and...

  2. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...physically joining multiple fragments of DNA to create a synthetically designed DNA sequence. There are...fragment joining, but this is fine for its goal of creating a choice of standardized constructs and tools ... regions. Golden Gate Assembly also allows the creation of standardized DNA constructs, but its use of... expediently join multiple synthetic fragments, create multiple mutations in one or multiple fragments...well-designed fragment overlap sections, while still creating scarless plasmid inserts.  Want to Learn More...and eliminates the annealing step usually used to create a double-stranded oligo. Generating an sgRNA construct...directly transformed into E.coli for plating, screening and generation of sgRNA. Generation of sgRNA libraries...

  3. Addgene Welcomes John Doench to the Advisory Board

    Type
    Blog Post
    Published
    Aug. 13, 2015, 2:30 p.m.
    ... primary screen to knowing what are the real hits is so much shorter…. it’s a great screening technology...come here, I need to spend time here, this is an incredible place.” Doench has spent most of his research...researchers in a variety of fields develop RNAi screens to answer their own research questions. Starting... For his post-doc, Doench worked on using RNAi screens to learn more about cancer biology in Ed Harlow...developing CRISPR pooled libraries for use in genetic screens and has contributed much of his knowledge to the...was completely justified… it has been just an incredibly useful technology.” He goes on to explain that...become more efficient in terms of editing and screening) such that thousands of individual edits can be...

  4. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...fusing miniTurboID with dCas9 to create TurboCas, the Shilatifard Lab created a very quick and efficient way...find in this post: Glutamate indicators with increased sensitivity and tailored deactivation rates Addgene's...targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation rates By...2025).In this latest work, the team designed and screened a large library of iGluSnFR3 mutants, identifying...Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation rates. bioRxiv...puromycin resistance gene makes stable cell line creation a breeze. Modular by design: Key elements (promoter...protocols for viral generation and stable cell line creation (Figure 2). Looking for more? Explore our empty...

  5. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    Published
    Dec. 21, 2021, 2:15 p.m.
    ...this approach can greatly increase the sensitivity of an assay by increasing the number of signaling molecules...green; and proteins are represented in orange. Created with BioRender.com.    How do secondary antibodies...shorter than that of the primary antibody, due to increased binding affinity, many indirect antibody protocols...are designed to take place over multiple days. Increased run time is considered one of the major drawbacks...specific isotype subclasses of primary antibodies can increase the possible number of target proteins.   The ...immunohistochemistry advances have found ways to increase signals from conjugated primary antibodies. The...as nanobodies, has introduced new options for increasing signal strength.  For the time being, though,...

  6. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ... the untagged allele. Created with BioRender.com. The first step in creating the plasmids was to identify...be prohibitively expensive for many labs. We’ve created and tested plasmids that use CRISPR/Cas9 to endogenously...first exon of the gene. In this manner, we have created an initial set of 10 plasmids representing 10 human...linker, and the GFP tag sequence to be inserted. Created with BioRender.com. A key feature of our methodology...the double strand break and introducing the tag. Created with BioRender.com. Plasmids, plasmids everywhere...strategy and design described above can be used to create similar donor plasmids for introducing tags into...

  7. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ... older plates could increase plasmid loss or mutation. Let it incubate Increasing the incubation time ...prep We'll discuss the following techniques to increase plasmid yield. Keep in mind that you might need...particularly stubborn prep. Check the bacterial strain Increase the volume Improve the culture media Let it incubate...more extensive listing of common E. coli strains. Increase the volume Starting with a larger culture volume...you are working with a low copy plasmid. When increasing the culture volume, be aware that a higher volume...origins of replication (e.g. p15A, ColE1) can be increased by adding chloramphenicol to the culture medium...glycerol or glucose (in moderation) may also serve to increase cell density and plasmid yield. Start with a single...

  8. Editor's Choice, October 2016

    Type
    Blog Post
    Published
    Nov. 4, 2016, 2:30 p.m.
    ...Laws Bring Increased Postdoc Wages Our most popular blog post this month shares incredibly important information...enacted labor laws will likely increase postdoc earnings in the U.S. This increase, while overdue in my opinion...considering a postdoc. In “Changing Labor Laws Bring Increased Postdoc Wages,” Future of Research executive director...variety of downstream effects including possibly decreasing the number of postdocs various academic institutions...Science Communication Changing Labor Laws Bring Increased Postdoc Wages I hope you all have a great November...

  9. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...experiment. Here, we will provide an overview of how to create and use a standard curve and provide some general... I make the standards for a standard curve? You create a standard curve by serially diluting a known control...mix the samples well by vortexing or inversion. Created with BioRender.com.   When choosing a standard... vortexing between each step of the series. To increase the accuracy of the curve, avoid pipetting small...value. How do I plot and use a standard curve? To create a standard curve, you graph the measured response...wider span of concentrations but suffers from decreased specificity, especially as it approaches the minimum... all of the samples in the assay, as this will increase the accuracy of the data. For assays involving...

  10. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox... [13]:  Germ cells [14] Hepatocytes [15] Pancreatic β-cells [16] Intestinal tissue [17] Lung epithelial...rare in mammals, an example is observed in the pancreas when excess β-cell damage results in the transdifferentiation...cell types such as fibroblasts, hepatocytes, and pancreatic exocrine cells have been successfully transdifferentiated...differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res, 2009. 19(4.... 38. Thorel, F., et al., Conversion of adult pancreatic alpha-cells to beta-cells after extreme beta-...PubMed PMID: 3690668. 40. Ferber, S., et al., Pancreatic and duodenal homeobox gene 1 induces expression...

  11. Hot Plasmids - November 2023

    Type
    Blog Post
    Published
    Nov. 14, 2023, 3:35 a.m.
    ...enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can increase on-target ...reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications.... The Ting lab recently made two deposits that increase the utility of this technique.  By engineering... O’Neill The Cohen lab and colleagues recently created a system for recording the transcriptional history...resembling a “ticker tape” recording. The authors created externally-timed bands of color with HaloTag-iPAK4...thin sections of tissue, the Wu Lab is working to create a three-dimensional atlas of protein expression...-95 [K28/43R] (70 µm mounted sagittal section). Credit: Zhuhao Wu, Weill Cornell Medicine.  Interested...

  12. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    Published
    Aug. 5, 2014, 6:51 p.m.
    ...Jim Woodgett. Public Library of Science (PLOS) created a stir earlier this year when it announced its ...manuscript submission. The rationale includes increasing the ability of reviewers (and, once published...some cases, typically consortia involving genomic screens, the data stream is so prolific that raw information...require access to each individual video. To PLOS’s credit, the organization subsequently clarified what the...and this alone is a reasonable motivation for increasing both data accessibility and methodological detail...Minimally, raw but sufficiently annotated data used to create charts and tables in the paper are in scope. For... up new opportunities for research as well as increasing transparency. Thank you to our guest blogger...

  13. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    Published
    March 22, 2016, 2:30 p.m.
    ... to each to make the library usable for pooled screens. Applications of the cancer pathways ORFs kit Wood...and his colleagues developed the kit for use in screening for pathways that drive resistance to drugs in...the only way to use the kit. “You could use it to screen for pathways that confer any phenotype of interest...drug resistance,” he said. “It could [be used to screen for] morphological features, migration, or survival...use our libraries to do things - sometimes drug screens and sometimes totally different experiments we ...Tracing Read How CRISPR Can Be Used in Genome-Wide Screening Browse 28 Hot Plasmid Technologies from 2015 ... cancer screening...

  14. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...Creating great educational science videos is tough. Is the content accurate? Are we conveying complex...people in tight locations. Conference rooms for on-screen presenters! Labs and benches! Everything was out...videos. It’s really nice to have a scientist on-screen, somebody who really knows the protocol and can...would be to use an iPad to shoot.  Build: Jennifer created a mobile camera area that included iPad and her...where I could focus on camera and audio, and the on-screen talent could just focus on their interview or performance...re-opened very early in the pandemic with some incredibly strict safety protocols to keep Addgenies safe...camera) would remain masked, it freed us up to be creative.  Test: Our first official shoot went off with...

  15. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...developing zebrafish pancreas. They first tagged these cells using plasmids. Then using HOTcre, a method that...of quiescent and proliferative pancreatic β-cells identified by HOTcre mediated labeling." Proceedings...these systems check out Gerald Rubin’s lab who created a set of modular vectors that can be used to optimally...unicellular organisms, they can also be used to easily screen multiple therapeutic compounds in one experiment...embryos with DNA or RNA to edit their genomes or to create transgenic animals. Zebrafish are an ideal model...β-cells that originate in different regions of the pancreas and produce different levels of insulin (Hesselson...

  16. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ...Symptoms of AIDS include rapid weight loss and an increased risk of infection, including both common infections...ART-treated patients lack plasma HIV-1, they are at increased risk for other chronic diseases, including dementia...disorders, nerve damage, and cardiac disease. This increased disease risk is attributed to latent HIV reservoirs... used histone deacetylase (HDAC) inhibitors to increase transcription of the integrated HIV-1 genome. ... enhancer region near the NF-KB binding sites increased activation of HIV promoter-luciferase constructs...microglial line, treatment with CRISPR/Cas9 SAM increased the percentage of cells with activated HIV-1, ...cell. ZFNs and CRISPR/Cas9 have also been used to create mutations in genes essential to HIV-1 replication...

  17. SunTag and Fluorescent Imaging

    Type
    Blog Post
    Published
    March 28, 2017, 2:30 p.m.
    ...scaffold. To increase expression, Tanenbaum et al. modified the GCN4 sequence to increase its alpha-helical...activation. Multiple nuclear localization signals (NLS) increase protein import into the nucleus. In developing... Weissman labs took this biological lesson and created a system to amplify fluorescent signals. Named ...protein is expressed at low levels, you have to increase your imaging time to get enough signal. This workaround...cells without affecting protein function, thereby creating a single-molecule reporter of intracellular processes...alpha-helical structure and stability, creating v4 SunTag. Since the v4 system does not display protein aggregation...

  18. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...resulting in decreased BRET2 signal. With increased concentration of drug, the BRET2 signal decreases. ...leads to downstream effects.  Bryan Roth’s lab created a collection of plasmids called TRUPATH to interrogate...the complex will disassociate, resulting in a decreased GFP2 emission. The activation state can be measured...showing the effectiveness of using TRUPATH as a screening technique. Lastly, the activity of both antagonists...plasmids are available as a collection, facilitating screening across the transducerome, to gain a more complete...

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

    Type
    Blog Post
    Published
    June 2, 2020, 1:15 p.m.
    ... Figure 1: Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  ... Peking University has developed a new CRISPR screening method that makes use of guide RNAs with internal...false-positive rate discovery rate, conventional CRISPR screens require a low multiplicity of infection (MOI) during... annotated human gene, it enables genome-wide screening with a high MOI, which is beneficial for a low... calcineurin dynamics and can serve as a drug-screening platform to identify new antifungals that target... assay, and are not suited to high throughput screening, calcineurin dynamics measurements, or monitoring... attached fluorescent protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry...

  20. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ...finding. Transcriptional program changes from this screen successfully differentiated cells resembling all...from Joung et al.)    Putting MORF to work Screens and selections In its current format, the MORF ...library is most immediately useful for TF library screens. Previously engineered TF libraries were not as... library can be used with other selection and screening mechanisms, although the unique barcodes for each...mutation and/or TF dysregulation. A targeted TF screening approach can not only lead to a better understanding...better simulate a disease in a model system. TF screens can be designed to identify custom TF programs ...Cas-9 knockout and transcriptional activation screening. Nat Protoc. 2017 (4):828-863. doi: 10.1038/nprot...
Showing: 181 - 200 of 798 results