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We narrowed to 813 results for: cell

Showing: 481 - 500 of 813 results

  2. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...there isn’t a template strand left behind that the cell can use to repair. Figure 3: General overview...complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394.e3. https://doi.org/10.1016/j.molcel...formation by a type I-C CRISPR Cascade. Molecular Cell, 83(5), 746-758.e5. https://doi.org/10.1016/j.molcel...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:...

  3. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ... 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 ...2016) Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins..., 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-...

  4. Plasmids 101: Gateway Cloning

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    Blog Post
    ...the next step is to transform competent E. coli cells and select the positive clones. The entry clone ... expression of the human KRAS gene in mammalian cells. STEP 1: Generate an Entry Clone There are a few...ccdB gene. Since the ccdB product is toxic to the cell, Gateway cloning efficiency can reach >99%. ...digest! Then, you can transform or transfect the cells that you want to use for your experiments and verify...Phage (Lambda) and Higher Organisms (Cambridge, MA: Cell Press). Additional Resources on the Addgene Blog...

  5. Antibiotic Resistance: An Old Solution but a New Problem

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    Blog Post
    ... attack the cell wall of bacterial cells; however, Mycoplasma species do not possess cell walls. As a ...bacteriophage (viruses that only infect bacterial cells), or a bacterium picking up resistance genes from...conserved, non-negotiable process within the bacterial cell, antibiotic resistance is very likely to develop...

  6. To Codon Optimize or Not: That is the Question

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    Blog Post
    ...mammalian cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this...sequence. That means there are 61 possible tRNAs. Cells however, may not express all 61 of these tRNAs and... are also optimized for expression in mammalian cells. For example, Henry Lester’s lab optimized the C...Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal...

  7. Chemogenetics Plasmids

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    Collection
    ...allowing a scientist to control signaling within cells expressing these engineered receptors. Chemogenetic...engineered receptor is coupled to and the specific cell type where the receptor is expressed, these receptors...

  8. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...your assay will scale. Or you don’t have enough cells to execute genome-wide perturbations. So, while ...lentiGuide expressing only an gRNA, for use with cells that already have Cas9). Each library consists of...be useful when screening for phenotypes, such as cell-non-autonomous effects, that do not lend themselves...

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

    Type
    Blog Post
    ...interaction networks of specific proteins in living cells. Until recently, the enzymes most frequently used...but requires H2O2, which can be toxic in living cells. BioID was derived from biotin ligase and uses far...Alexander Green lab found a solution: a low-cost, cell-free detection assay that can be completed without...family, known to be involved in cytoskeletal control, cell growth, and endocytosis. Recently, the Sidhu group...to studying membranes, organelles, and other subcellular compartments is via proteomics--the study of ...

  10. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ... R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Screen identifies... R, Schmalhorst PS, Jude JG, Zuber J, Gerlich DW Cell. 2017 Aug 24;170(5):956-972.e23 Rapid and phosphoinositol-dependent... Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural role..., Ensminger AW, Fox AH, Chess A, Lawrence JB Mol Cell. 2009 Mar 27;33(6):717-26. Reporter-nanobody fusions...

  11. Troubleshooting Your Plasmid Cloning Experiment

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    Blog Post
    ...product, the transformation may not result in any cells, or all the colonies screened might not contain ...verify the strain and the efficacy of your competent cells (when they get old, they lose competency). If all... look at your experimental design. Full lawn of cells. The antibiotic in your plate might not work. Check...acknowledged in hundreds of publications including Cell, Nature, and other journals.   Paolo Colombi is ...

  12. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...also gives cells antibiotic resistance to neomycin so that you can select the population of cells with the...Only a small percentage of the proteins inside a cell have a commercial antibody against them. There’s...antibody. It’s an epitope not found in human or other cells. It was made artificially. That means you aren’t...

  13. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...successfully infected/expressing cells by flow cytometry. The isolated cells were injected into mice and imaged...to changes in intracellular calcium and is useful for direct imaging of events in cells and brain explants...packaged into a retrovirus for infection of host cells. The fluorescent moiety of the BRET reporter was...imaging multiple proteins at the same time in live cells using fluorescent proteins (FPs) can be problematic...available through Addgene (Plasmid #83926). Live-Cell Voltage Sensor  Ingaki and colleagues (8) were interested...including monitoring multiple cellular events such as the dynamics of subcellular structures and gene expression...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors...

  14. Rinehart Lab Phosphoprotein Reagents

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    Collection
    ...abundant forms of posttranslational modifications in cells and research into its many roles in protein function...interactions can be discovered by fluorescence-activated cell sorting (FACS). The mode #2 phosphosite library ... Production of Recombinant Phosphoproteins Using Cell-Free Protein Synthesis. Oza JP, Aerni HR, Pirman...Ordureau A, Paulo JA, Rinehart J, Harper JW. Molecular Cell . 2015. Sep 9. pii: S1097-2765(15)00662-0. doi: ...

  15. The 12 Days of CRISPR

    Type
    Blog Post
    ...function using transcriptional recording sentinel cells! 🎵 On the 11th day of CRISPR, Addgene gave to me...activation and interference screens for primary human T cells🎵 On the 3rd day of CRISPR, Addgene gave to me: ...Enhanced prime editing systems by manipulating cellular determinants of editing outcomes🎵 On the 5th ...

  16. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...repair fix DNA double-strand breaks?  In eukaryotic cells, repair of DSBs occurs primarily by two pathways...used, however, these are generally more toxic to cells than their plasmid counterparts. Another consideration...how to fix a broken relationship. Pardo B, et al. Cell Mol Life Sci (2009). PubMed. Regulation of DNA...break repair pathway choice. Shrivastav M, et al Cell Research (2008). PubMed. Mechanisms of DNA double-strand...Improve HDR Efficiency for CRISPR Editing in Human Cells Resources on Addgene.org Browse All CRISPR Plasmids...double-strand break (DSB). DSBs can be caused by intracellular factors such as nucleases and reactive oxygen...

  17. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.org/10.1016/j.cell.2015.09.038 Find plasmids...are capable of cleaving genomic DNA in mammalian cells and zebrafish embryos, and they can be used to modify... adapted for targeted RNA cleavage in mammalian cells. The REPAIR (RNA Editing for Programmable A to I...reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol 34:863–868 . https://doi.org/10.1038...specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol 34:869–874 . https://doi.org/10.1038...

  18. Antibodies 101: Conjugation

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    ...and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org... like proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et...epitope and transcriptome measurement in single cells. Nat Methods 14:865–868. https://doi.org/10.1038...

  19. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing).  Some...enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol 33, 985–989 (2015). https://doi....Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021). https:/...deactivation with photocleavable gRNAs. Molecular Cell 81, 7, 1553-1565 (2021). https://doi.org/10.1016...evolved ways to prevent degradation of important cellular RNA molecules with post translational modifications...transfection efficiency and even track localization in cellular experiments. There are many suitable dyes to choose...

  20. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ... be expressed in the same cell, but not necessarily at the same subcellular compartment as the opsin or...modulate the activity of neurons and other brain cells. Since the opsins or chemogenetic receptors used...neuropil conceals identification of opsin-expressing cells). For other things to keep in mind when making fluorescent...cerevisiae by use of an epitope addition method. Mol Cell Biol 8:2159–2165 . https://doi.org/10.1128/mcb.8.5.2159...
Showing: 481 - 500 of 813 results