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We narrowed to 813 results for: cell

Showing: 421 - 450 of 813 results
  1. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

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    Blog Post
    ...expressing target cells. Any neutralized pseudoviruses would be unable to get into the target cells and this can...disease severity and survival. Cell 184:476-488.e11. https://doi.org/10.1016/j.cell.2020.12.015 Garcia-Beltran...vaccine-induced humoral immunity. Cell 184:2372-2383.e9. https://doi.org/10.1016/j.cell.2021.03.013 Additional resources...vaccines and allows the virus to gain entry into human cells via human ACE2 receptors. In their assay, the pseudoviruses...
  2. Luminescent Imaging with Nano-lanterns

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    ...have become indispensable tools for molecular and cell biologists over the last two decades, but their ...quantum yield, and when transiently expressed in cells, generates much less signal than comparable fluorescent...three luminescent proteins can be used together in cells. As Nano-lanterns are not subject to photobleaching...Nano-lanterns, Takai et al. continuously imaged cells for about 4 hours! The latest enhanced Nano-lanterns... al. “Luminescent proteins for high-speed single-cell and whole-body imaging.” Nat Commun. 3 (2012): 1262...light. Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be...phototoxicity, they successfully imaged multiple subcellular compartments over several minutes using Nano-...
  3. CRISPR 101: Epigenetics and Editing the Epigenome

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    ...mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced cells. Grant...in the Mammalian Genome. Cell 167:233–247.e17 . https://doi.org/10.1016/j.cell.2016.08.056 Find plasmids...targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids from...for targeted cytosine demethylation in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD ...vectors and targeted delivery into hard-to-manipulate cell types. They have also fused LSD1 to x-Cas9(3.7)....modifiers may be more frequently inherited by daughter cells. For example, KRAB induced silencing was transient...enhancers. As the effects of these tools are likely cell type- and context-dependent, it may make sense to...
  4. Recombinase-based State Machines Enable Order-dependent Logic in vivo

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    ... require minimal cellular perturbation. Also, this method requires live, intact cells; the sequencing ...Synthetic recombinase-based state machines in living cells” (1), into a more straight-forward, concise explanation... our paper (1), we allowed individual bacterial cells to form colonies on a plate and then we performed..., but more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The...sufficient for greater than 50 percent of RSM-containing cells in a population to correctly record each possible... were sufficient for greater than 90 percent of cells to correctly record each possible permuted substring...Synthetic recombinase-based state machines in living cells. Science. 353, aad8559 (2016). PubMed PMID: 27463678...
  5. Six Spooky Science Stories and Halloween at Addgene

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    Blog Post
    ...attaches to the cell wall of green algae (Soo et al., 2015). There, it divides into two cells. At the same...Vampirovibrio). What’s left of the algae cell is just the cell wall and cytoplasmic membrane. Vampirovibrio...biosynthesis in Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID...
  6. Split Fluorescent Proteins for Studying Protein-Protein Interactions

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    Blog Post
    ...measuring protein expression or localization in cells, or even single-particle tracking of the bound complex...(1–10) and GFP(11) Versatile protein tagging in cells with split fluorescent protein. Kamiyama et al. ...tracking of protein-protein interactions in living cells. Shao et al. Commun Biol. 2021 Yujie Sun   For...bright enough for single-molecule tracking in live cells.  Controls  When designing experiments, it’s important... fragments themselves can sometimes interact in cells and produce background signal (especially for some...some older split-GFP constructs or certain cell environments). Also consider that a protein fusion may...) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent protein tags. ...
  7. Of Myc and Men

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    ...For about as long as the disciplines have existed, cell and molecular biologists have been continuously ... peer into the mysterious, microscopic world of cells. The development of antibodies as tools allowed ...monoclonals would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different...specific for human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616. https://doi.org/10.1128/mcb.5.12.3610... and immunoglobulin heavy chain binding protein. Cell 46:291–300. https://doi.org/10.1016/0092-8674(86...signal prevents secretion of luminal ER proteins. Cell 48:899–907. https://doi.org/10.1016/0092-8674(87...
  8. CRISPR Cheat Sheet

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    Blog Post
    ...nuclease target multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of...range. CRISPR can be difficult to deliver. Not all cells efficiently take up plasmids used to produce CRISPR...
  9. Antibody Data Repositories and Search Engines

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    Blog Post
    ... Immune cells Flow cytometry (potentially others) Data repository From the Human Cell Differentiation...repository   HuBMAP Consortium Healthy human cells Imaging (IHC, ICC, IF, etc.)  Data repository...
  10. New Viral Vectors - Fall 2024

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    Blog Post
    ...the fall colors are inspiring you to paint your cells in equally vivid colors, you'll want to check out...AAV9-X1.1 enables expression in brain endothelial cells via an injection into the systemic circulation. ...levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also...recombinase or a fluorescent marker in the various cell types of the basal ganglia. They work via newly-...
  11. Antibodies 101: Polyclonal Antibodies

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    ...system, antibodies are produced by B cells. Each individual B cell produces antibodies that all recognize...also all the same isotype. But together, all the B cell clones in the immune system make different isotypes...antibodies starts with selecting just one of these B cell clones for further antibody production, polyclonal... blood from the immunized animal. The red blood cells and the serum are separated, and the serum - which...
  12. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

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    ...editor in mammalian cells Not all Cas13d proteins are functional in mammalian cells, but Konermann et al...make it possible to target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding... with RNA-Targeting Type VI-D CRISPR Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: ... a WYL-Domain-Containing Accessory Protein.” Mol Cell. (2018) pii: S1097-2765(18)30173-4. PubMed PMID:...
  13. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    ...Drosophila cell line, I’ve studied a transcription factor important for T cell differentiation (a cell type ...regeneration and homeostasis in the Drosophila midgut. Cell. 137, 1343–55 (2009). PubMed PMID: 19563763. PubMed...et al. Digoxin and its derivatives suppress TH17 cell differentiation by antagonizing RORγt activity. ...
  14. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

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    ...indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured. Archon1 ... it. Using a robotic picking approach to select cells with variant characteristics they wanted, the team...approach to control protein-protein interactions in cells because the protein binders can be customized for...TOM20 (pTriEX Tom20 mVenus BoPD). In dark adapted cells expressing these plasmids, tgRFP fluorescence localizes...offers a way to spatially and temporally control cellular processes in vivo. Many photoactive proteins exist...photo-reversible protein-protein interactions can control subcellular localization processes in vivo. They deposited...
  15. Fluorescent Proteins 101: Luciferases

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    ...luciferase assays require cell lysis as the most efficient means to disrupt the cell membrane and deliver ...produced. A similar approach can be used to analyze cell signaling pathways or miRNA knockdown. BRET for ...advantageous properties. This approach can improve live-cell imaging by avoiding autofluorescence and phototoxicity...can permit measurements of luminescence from live cells. Luciferase variants with incompatible substrates.../phototoxicity Not susceptible Susceptible Subcellular imaging Improving Well-established High-throughput...
  16. R Bodies: Membrane-Rupturing Microscopic Tools

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    ... can rupture E. coli spheroplasts (cells which have had their cell walls removed with lysozyme). This ...protein fusions allowed us to image R bodies inside cells, and these constructs may be useful to future scientists...applications. For example, you could use them to lyse cells (and surrounding membranes) in keeping with their...assemble and function, we might be able to endow cells with new ways to interact mechanically with their...penetrate other barriers (like vasculature or extracellular matrices) in response to pH changes. Beyond...
  17. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    ...Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;156(5):935-49. doi: 10.1016/j.cell.2014.02.001. Epub 2014 Feb...Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct...screening for genes that, when turned on, make melanoma cells resistant to treatment with a BRAF inhibitor. (The...Weissman and his colleagues demonstrated in a paper in Cell that they could typically achieve 90-99% knockdown...straightforward to apply the new library for use in various cell types and in live mice. Therapeutic uses in humans...Addgene, enabling systematic investigation of the cellular consequences of repressing or inducing individual...
  18. Neuronal labeling with Spaghetti Monster

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    ...ability to interact with protein partners within the cell. However, antibodies can have low affinity for these... generally bright, stable and well tolerated by cells. They can be used for protein localization, isolation...trigger protein aggregation which is toxic for the cells and affects the labeling of small structures like...team, an ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with...example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays a critical role in neural...Advances in fluorescent protein technology." Journal of cell science 120.24 (2007): 4247-4260. PubMed PMID: 18057027...of neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in the...
  19. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

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    ...Proteins in Living Cells Using Bimolecular Fluorescence Complementation. Molecular Cell 9:789–798. https...manner. This is straightforward in easy-to-transfect cell lines where plasmid dosage can be tightly controlled...fluorescence-based method for directly visualizing PPIs in cells. The ease with which BiFC can be visualized makes...visualization of protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038...(BiFC) Analysis of Protein Interactions in Live Cells. Cold Spring Harb Protoc 2013:pdb.prot076497. https...directly visualize PPIs that regulate numerous cellular functions, including ubiquitinoylation, kinase...BiFC studies used the method to interrogate the subcellular localization of interactions between basic leucine...
  20. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    ... systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016/j.cell.2023.03.034 Resources...Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016/J.CELL.2013.07.018 Anzalone, A....epigenetic manipulation of prime editing. Cell. https://doi.org/10.1016/j.cell.2024.03.020 Mathis, N., Allam, A...suited for prediction of very diverse edit types and cell contexts. Figure 1: High-throughput screening...double-strand break repair pathway balance. Molecular Cell, 81(10), 2216-2230.e10. https://doi.org/10.1016/...
  21. Fluorescent Proteins 101: Aptamer Fluorophores

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    ..., peptides, amino acids, drugs, metal ions, and cells (even pathogenic bacteria). That range is possible... combination of SELEX and fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer...and FACS, Broccoli is inherently more stable in cells. The new aptamer also has a higher melting temperature...cassettes, thereby extending their use into mammalian cells. They’ve also dimerized Broccoli to further enhance...Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. USA 112(10) (2015): 3002...Aptamers as a Tool to Study RNA-Modifying Enzymes.” Cell Chem Biol. 2016 Mar 17;23(3):415-25. PubMed PMID... over half a century; the Addgene blog has an excellent primer and tips from experts on FRET techniques...
  22. Fluorescent Proteins: FRET

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    ...influenced by many factors, such as the organism or cell type, relative position and orientation of the fluorophores... (Unpublished) FRET Indicators - Live and Fixed Cells Additional Resources at Addgene Learn more on the...
  23. Addgene’s Top 10 Blog Posts from 2014

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    ...more about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating many additional... We’re happy to see scientists making their own cells to save money and create the tools that work best...Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids 101: The Promoter Region...
  24. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

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    ...Multiplex genome engineering demonstrated in human cells They used the Golden Gate cloning method to ligate...successfully to target multiple sites in cultured human cells. While it has not yet been demonstrated, they say...Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sakuma...
  25. CRISPR 101: Off-Target Effects

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    ...millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to sequence...trying to generate a knockout cell line, with the goal of isolating single cell clones to use for multiple...generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation arising.... a level of, say 5%, then the majority of single cell editing events will be on target. Thus, if you isolate...example, if your goal is to generate a knock-out cell line to perform downstream characterization of, ...
  26. Evolution of Lab Techniques

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    ...Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically advanced...gene mutation is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with... of the mutated β globin gene that causes sickle cell anemia. The mutation happens to destroy a sequence...conducted on a sample from an individual without sickle cell anemia, two DNA fragments are observed because the... site where MstII cuts. In a patient with sickle cell anemia the enzyme does not cut the DNA so only one...Wai Kan. "A sensitive new prenatal test for sickle-cell anemia." New England Journal of Medicine 307.1 (...transgenic constructs and mice." Current Protocols in Cell Biology (2009): 19-10. PubMed PMID: 19283728. PubMed...
  27. Viral Vectors 101: Optogenetic Tools

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    ...pulses to the subject/cells expressing the optogenetic tool causing a change in cellular activity or gene ...positively charged ions to rapidly pass through the cell membrane and initiate an action potential - a very...such as Na+, K+, and Ca++ to passively enter the cell with the concentration gradient to depolarize it...the concentration gradient and hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine...different wavelength, turns it off, allowing the cell to be excited over a sustained period of time (Gong...affect how rapidly you can activate or inhibit a cell in succession and maintain fidelity with the stimulus...many downstream signaling mechanisms that are often cell-type specific. If it is something you’re considering...
  28. CRISPR 101: Cytosine and Adenine Base Editors

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    ...that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using the U-containing strand..., which may lead to cell death rather than NHEJ repair, thus removing these cells from the edited population...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual base...relatively low frequency, especially in nondividing cells. Scientists everywhere wistfully imagined being ...editing efficiency, Komor and team wanted to force the cell to use the deaminated DNA strand as a template. ...frequency to above 30% for a variety of targets in human cell lines, with an average indel frequency of only 1.1%...modifications. CRISPR base editing persists through multiple cell divisions, indicating that this method produces ...
  29. Human Germline Editing Using CRISPR

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    ...ssODN repair template in induced pluripotent stem cells made from the patient. With the first gRNA, they...homology-directed repair (HDR) taking place in 41.2% of edited cells. The results were even lower for the second gRNA...13.1% targeting efficiency with only 13% of edited cells employing HDR. Ma et al. moved forward with gRNA...babies.” The differences in editing between iPS cells and embryos make a case for conducting research ...analysis is not possible with IVF, as most of the cells are needed for proper development. Ma et al.’s experiments...as whole exome sequencing of embryo derived-ES cells.They also conducted whole genome sequencing in selected...
  30. CRISPR 101: Cas9 vs. The Other Cas(s)

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    ...Deletions in Human Embryonic Stem Cells Using Type I CRISPR-Cas. Molecular Cell, 74(5): 936-950.e5 (2019). 10.1016...a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016/j.cell.2015.09.038 Harrington, B., ... Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral RNA...cleavage and is significantly less toxic to mammalian cells than Cas13 (likely due to the lack of collateral...
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