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We narrowed to 872 results for: cel

Showing: 481 - 500 of 872 results
  1. Viral Vectors 101: Virus Safety

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    ...producer cell line, such as HEK-293T, production of viral particles, and transduction of a target cell line...re derived from viruses that can infect people, cells, and animals. Some of these viruses are completely...While the virus can still enter (and thus infect) cells, this safety feature prevents the virus from replicating...replicating and spreading in non-target cells, including yourself as the researcher.   Figure 1:...vector carries an shRNA to a gene essential for cellular function, this would also be a higher risk vector... the procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-...
  2. Illuminating Epigenetics with A FRET Based Biosensor

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    ...genes should be ‘read’ by the cellular machinery. This is how most cells in the body contain the same ...complex of DNA and histone proteins that compacts cellular DNA into chromosomes and reduces the risk of dangerous...the more compacted it becomes. This prevents the cellular machinery from physically interacting with the...visualise changes in histone methylation in living cells. Essentially this reporter construct consists of...recognise lysine 27 methylation. When tested in living cells where methylation of these sites was controlled,...fluorescent reporters of histone methylation in living cells." Journal of the American Chemical Society 126.19...
  3. Degrading DNA with Cascade-Cas3

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    ...unwinding steps. Molecular Cell, 70(3), 385-394.e3. https://doi.org/10.1016/j.molcel.2018.03.031 Xiao, Y., ...CRISPR Cascade. Molecular Cell, 83(5), 746-758.e5. https://doi.org/10.1016/j.molcel.2023.01.024 Morisaka, ...there isn’t a template strand left behind that the cell can use to repair. Figure 3: General overview...broad and unidirectional genome editing in human cells. Nature Communications, 10(1). https://doi.org/10.1038...CRISPR–Cas systems for genome engineering in human cells. Nature Biotechnology, 37(12), 1471–1477. https:...
  4. Getting to Know Expansion Microscopy

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    ... for EM is not something you find in the average cell and molecular biology lab, and EM is limited in ...ultrastructural relationships that influence everything from cell division to memory, but don’t have access to the...fix (and maybe label) your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample...properties of the sample itself. For example, cultured cells are typically easier to expand than mouse brain ..., Mohar B, Eddison M, et al (2022) Visualizing cellular and tissue ultrastructure using Ten-fold Robust... Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures using expansion microscopy (U-...2016) Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins...
  5. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    ...Generating a no-fuss genetic knockout  Knock-out cells or animals are engineered by introducing Cas9-mediated...transcription by blocking its initiation. In mammalian cells, robust repression requires targeting dCas9 along...study essential genes whose knockout would cause cell lethality. The Krüppel associated box (KRAB) is ...repressing target genes in a variety of species and cell types can be found on Addgene’s website. Activating...activating target genes in a variety of species and cell types can be found on Addgene’s website. ...optimized for expression in different species and cell types including (but not limited to) bacteria, yeast... have delivered Cas9 and a sgRNA to your target cells, it is time to confirm that your target sequence...
  6. CRISPR 101: Any Base Transversion Editors

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    ...base editing. Molecular Cell, 84(7), 1257-1270.e6. https://doi.org/10.1016/j.molcel.2024.01.021 Tong, H.,... edit A → Y (Y = T or C) in 67.17% of screening cells (Tong, Wang, et al., 2023). Based on this promising...the base editor. Black arrows indicate natural cellular processes. Created with BioRender.com. The...Cas9 nicks the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template... efficient A•T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4), 638–650...efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547. https:...
  7. A Scientist's Guide to Ebola

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    ... infected cells to go undetected by the immune system in a novel way, by shielding host cell surface proteins... itself to escape vesicles within host cells and turn the cell into a veritable virus factory.  Their ... sets off.  The virus works its way into immune cells in ways that aren’t fully understood. Much of the... viral attachment and fusion. Once inside a host cell, new evidence in PLoS Pathogens shows that the Ebola...
  8. Plasmid Preps: Different Purity, Different Quantities, Different Uses

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    ...lipopolysaccharide of the cell wall of Gram negative bacteria like E. coli that affects tissue culture cell line viability...introduces transformation grade plasmids into a bacterial cell. Transformation grade DNA  Transformation is ...transformation of chemically and electrocompetent cells is minimal (picogram range). It can be extracted...Figure 3: Need to introduce plasmids into mammalian cells? Use transfection grade DNA. Transfection is ...process of inserting an external plasmid in mammalian cells and it will likely require micrograms of plasmid...
  9. Antibodies 101: Introduction to Immunofluorescence

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    ...interest live within a cell,” “Does this disease change the architecture of my cells,” or “How does this ...IF), is an immunoassay that brings to light the cellular world. The technique allows you to ask questions...this mutation impact the types of cells found in my tissue.” It is based on the same principles as other ... visualize a target of interest within an intact cell, tissue, or, in some cases, a whole organism. But...joined to fluorescent molecules are applied to a cell or tissue sample where they bind to their targets...to visualize a target within an intact sample, a cell in the case of ICC or a tissue in the case of IHC...
  10. Plasmids 101: Expressing shRNAs for RNA Interference

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    ...silencing complex. Molecular Cell, 82(1), 30–43. https://doi.org/10.1016/j.molcel.2021.11.026 Kilikevicius...miRNA and piRNA can be found endogenously in many cell types, while shRNA and siRNA are synthetic constructs...methods. shRNA processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the...shRNAs is to generate stable knockout or knockdown cell lines. In fact, it was the go-to choice for many...shRNAs are a quick way to create stable knockdown cells lines, and you can usually find validated shRNA ...Resources References Bartel, D. P. (2004). MicroRNAs. Cell, 116(2), 281–297. https://doi.org/10.1016/s0092-...
  11. 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...
  12. Supporting Rare Disease Research with BeHeard 2017

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    ...partners in mouse cell lines. Dr. Di Schiavi hopes that these studies will accelerate the development of...learn how plasmids from Addgene will be used to accelerate rare disease research and provide hope to patients...plasmids to upregulate their expression in specific cells. With a more concrete understanding of the molecular... of ~30 patient derived peritoneal mesothelioma cells lines and assess their tumorigenesis. These experiments...effects on the disease. This work will hopefully accelerate the development of targeted therapy for peritoneal...
  13. 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, ...
  14. Plasmids 101: Dimers and Multimers

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    ...at lower copy numbers within bacterial cells, and bacterial cells with multimers grow more slowly compared...stable and more prone to being lost in bacterial cells compared to monomers, although this can be prevented...increase the frequency of base substitution in human cells. J Biol Chem. 2011 Mar 25;286(12):10017-26. doi:...plasmids leading to the formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674...supercoiled plasmid DNA concatemers in mammalian cells. J Gene Med. 2009 May;11(5):444-53. doi: 10.1002...essential for plasmid monomerization and stability. Cell. 1984 Apr;36(4):1097-103. doi: 10.1016/0092-8674...
  15. DAP arrays for multiplex gene editing

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    ...different types of human cells, including T cells and hematopoietic stem and progenitor cells. Now with the DAP...to enable multiplex gene editing in a variety of cells and organisms. However, lengthy genetic payloads... over 50% averaging editing efficiency in human cells. MBE with DAP array also exhibited reduced Cas9-... editing, and nuclease editing, in virtually any cell type or organisms of interest with only minimal ...
  16. Scientists Map the SARS-CoV-2-Human Interaction Network

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    ... used HEK293T cells in our first study, but we're planning to soon expand to other cell types, including...26 of the 29 SARS-CoV-2 viral proteins in human cells and used these viral “bait” proteins to fish for...human proteins that the virus interacts with in cells. Because viruses rely on the help of human proteins...monkeywrench into the virus’ hijacking of human cells. Even better, more than half the drugs hitting these...they have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a fairly...
  17. K. phaffii: Rising to the Occasion

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    ...path.   Yeast as a model organism Yeast are single cell, eukaryotic organisms formally classified as fungi...and S. cerevisiae (budding). Yeast make for an excellent model organism for a few reasons: Affordable...similarity to the ancient yeast strains and metazoan cells.   Applications for K. phaffii There’s a number ...protein bang for your media buck - it grows at higher cell densities than most of its counterparts, yielding...oxidative reactions. They are found in all eukaryotic cells and peroxisomes are where methanol metabolism occurs...in methylotrophic organisms. K. phaffii is an excellent model for peroxisome studies since their peroxisomes...
  18. 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...penetrate other barriers (like vasculature or extracellular matrices) in response to pH changes. Beyond...assemble and function, we might be able to endow cells with new ways to interact mechanically with their...
  19. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    ...gives a talk at the Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and Beyond meeting...III product is produced and can infect bacterial cells. If a phage does not deliver an active variant, ...passage from the infected to a fresh uninfected host cell culture). This dilution step represents the major...but gets manually passaged from an infected host-cell culture to an uninfected culture which results in...
  20. 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 .../phototoxicity Not susceptible Susceptible Subcellular imaging Improving Well-established High-throughput...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...
Showing: 481 - 500 of 872 results