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We narrowed to 813 results for: cell;

Showing: 181 - 200 of 813 results

  1. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...numerous cell biology assays. However, there has been a pervasive view that most FPs in a cell will fold...fluorescent fusion proteins in cell biology."Current Protocols in Cell Biology (2005): 21-4. Pubmed PMID...Lee. "Fluorescent proteins: a cell biologist's user guide." Trends in cell biology 19.11 (2009): 649-655...Fluorescent proteins in cellular organelles: serious pitfalls and some solutions." DNA and cell biology 32.11 (...-fusions on quantitative imaging experiments and cell physiology in general. Pitfalls of Fluorescent Protein...quantitative imaging experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable...dynamics of fluorescent protein fusions in live cells in real time. Previously, we have described a number...

  2. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ... trigger the activation of dendritic cells, macrophages, T cells, and the recruitment of neutrophils. ... says. In induced pluripotent stem cells derived endothelial cells from women who are missing an entire...detect molecules outside of the cell and translate those signals into cellular responses. For example, GPCRs...maintaining the nutrient and ionic balance of the cells,” says Dr Sangeetha Iyer, ACD’s Scientific Advisor...that solute channels typically expressed on other cell types can compensate for or bypass the dysfunction...collection to perform a functional screen in mammalian cells to identify any solute channels that can compensate...be able to validate their findings in mammalian cells with both human and murine targets thus informing...

  3. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    ...on freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free extracts Typically... into a host cell (through a process called transformation), essentially giving the cell all the instructions...to dispose of the live cells. The alternative is to express the protein in a cell-free manner. This is ...electrophoresis to separate DNA or transforming cells with green fluorescent protein, these activities...transcription and translation. Unfortunately, having live cells in a classroom presents equipment needs, such as...operating the rest of the car as well. Similarly, cell free expression systems contain all the parts needed...handle a living, replicating organism. Furthermore, cell free systems can be freeze-dried into pellets (FD-CF...

  4. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ... where many different cell types, or even different phenotypes of the same cell type, are intermingled...postsynaptic cells when used at very high titers (Zingg et al., 2017), which can lead to cells in far away...Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org/10.3390/cells12050785 Murlidharan, G., Samulski...Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org/10.3390/cells12050785 Kanaan, N. M., Sellnow...experimental conditions and measurements; (2) the cells you’re targeting; (3) how much expression is needed... capsids, which determine how a virus enters the cell. Different serotypes utilize a number of different...co-receptors, and other mechanisms to gain entry into the cell (Dudek et al., 2018; Korneyenkov & Zamyatnin, 2021...

  5. An Introduction to Adenovirus

    Type
    Blog Post
    ... AdVs is dependent on modulation of the host cell’s cell cycle; inhibition of the innate immune system... in the producer cells where there is E1 and E3 gene complementation. In target cells that are infected...death of adenoid cells in vitro (Rowe et al., 1953). While overt death of the targeted cells is less likely...infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(4), 2296.... & Verma, I. (1998). Cellular immune response to adenoviral vector infected cells does not require de ...production and/or by genes present in the HEK293T cell line used (which have fragments of the AdV genome...achieves persistent transgene expression in certain cell types (Tatsis et al., 2007), potentially because...

  6. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...measure, and control molecular signals, cells, and groups of cells in order to understand their activity...Fluorescence-activated cell sorting (FACS): This is a type of flow cytometry that separates mixtures of cells into distinct...Thus, FACS can be used to separate cells expressing GFP from cells that are not. Developmental/transgenic... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...monitor gene expression from that promoter in a given cell type. This type of transcription reporting was among...find optogeneic actuators and sensors at Addgene. Cell marking/selection: Expression constructs like plasmids...

  7. AAV Molecular Tools

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    Collection
    ...Scanziani Tools for Cell Ablation These AAV encode tools that can be used for cell ablation. ID Name Expression...Transactivators Affinity Purification Neurophysiology Cell Ablation Tetracycline Transactivators and Inducible...diphtheria toxin receptor fused to GFP for studying cell ablation. 2 Jessell , Azim 45580 pAAV-flex-taCasp3...expression of designer pro-taCasp3 and TEVp for studying cell ablation. 5 Shah , Wells Don’t See What You’re Looking...

  8. Zinc Finger Consortium: OPEN Reagents

    Type
    Collection
    ...function in zebrafish, plants, and human cells ( Maeder et al., Mol Cell 2008 , Foley & Yeh et al., PLoS ONE... Townsend et al., Nature 2009 , Zou et al., Cell Stem Cell 2009 ). Resources A detailed protocol for practicing...ZFNs that function in zebrafish, plants, and human cells Kits...

  9. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...difficult-to-transfect cell types ranging from primary to embryonic stem cells. Viral integration of a... also generally less toxic to cells than transfections, preserving cell viability. Library delivery Screening...Viral applications Viruses work by infecting a host cell (the target of viral infection) and delivering a...to be anything you want delivered to your target cells. This delivered DNA/RNA can then be either permanently...permanently integrated into the genome of the target cell or transiently expressed. It’s important to note...This vector can express your construct in target cells transiently or constitutively depending on the virus...transcription factors, barcoded libraries to trace cell lineage, and so much more! Libraries allow you to...

  10. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    ...4,000 cell lines. Seventeen of these cell lines were used to investigate the susceptibility of cells from...additional cell line used in this study came from the Japanese Collection of Research Bioresources Cell Bank...that SARS-CoV, which entered human cells via hACE2, could not enter cells expressing murine ACE2 (Subbarao...repositories that distribute mouse strains, plasmids, cell lines, and much more have been critical for pandemic...from Reproducibility for Everyone. Case study 1: Cell lines Many of the reagents used to investigate SARS-CoV...accelerating research into this virus. One such example is cell lines. The American Type Culture Collection (ATCC...of this pathogen. The sharing and distribution of cell lines from repositories on opposite sides of the...

  11. Hot Plasmids - May 2022

    Type
    Blog Post
    .... Nat Cell Biol. 2022. https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal...with one visual barcode per cell and up to 200-plex with two barcodes per cell. The researchers validated...long sequence knock-ins Visual barcodes for live cell clonal multiplexing Improving flexibility for ... powerful approach for proteomic mapping in both cell culture and whole organisms (see our 2019 overview...reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally have higher risks ...multiplexing by: Rachel Leeson Multiplexed images in live cells remains a challenge due to multiple factors, including...flourescent proteins that can be inserted into a cell without disrupting its functions. Recently, Ravid...

  12. Allen Institute for Brain Science AAV Enhancer Collection

    Type
    Collection
    ...to target a variety of cell populations in different brain regions. Labeled cell populations were tested...regulatory sequences — termed enhancers — to drive cell type-specific expression. The Allen Institute for...differential chromatin accessibility across different cell types and cloned into an AAV backbone. Most enhancers...mouse brains. This collection contains plasmids for cell populations in the following regions: Isocortex ...-WPRE3-BGHpA AiP12320 AiE0475m SYFP2 Chandelier cells (Pvalb) Isocortex 230384* pAAV-AiE0475m-minBG-iCre...pAAV-AiE0475m-minBG-iCre(R297T)-BGHpA AiP13790 AiE0475m Cre Chandelier cells (Pvalb) Isocortex 214561 pAAV-AiE2566m-minBG-SYFP2...pAAV-AiE2150m-minBG-SYFP2-WPRE3-BGHpA AiP1379 AiE2150m SYFP2 Purkinje cells Cerebellum 208141 pAAV-AiE0390m_3xC2-minBG-SYFP2...

  13. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...fluorescence-activated cell sorting (FACS) helps researchers sift through thousands of cells to identify cells exhibiting...target single cells. Now, Francois St-Pierre’s lab developed a new form of FACS-based cell screening, Single-cell...individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures.  HeLa cells expressed... conformational changes in a protein.    Cells expressing an RFP reporter plasmid for recombinase...is active in vitro as well as in human and plant cells. Casɸ from Biggiephage can eliminate competing... Single-cell Phenotypic Observation and Tagging with Light (SPOTlight) as a way to isolate individual ...of keratin with mGold (top) and mVenus (bottom). Cells were imaged every 30 seconds for 3 hours. Image ...

  14. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ... on the method used to introduce the gRNAs into cells. Most CRISPR pooled libraries have been created ...the chance of transducing a only a single gRNA per cell. The viral DNA containing the gRNA then integrates...next-generation sequencing of the genomic DNA. However, some cell types are poorly transduced by lentivirus, and thus...CRISPR libraries was not possible. Primary murine T cells cannot be transduced by lentivirus and have been...library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express Cas9 and screen...CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction, ..., now enabling forward genetic screens in cells with few genetic tools. Find the mouse CRISPR knockout...

  15. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer, Inflammation and Immunity and...recent findings in basic immunology, cancer-immune cell interactions, cancer immunotherapy, as well as new...example the upregulation of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment...follow the interactions between tumor and immune cells in real time. Addgene depositor Professor Max Krummel...effect of PD-1 blockade.“ (Ref 4). PD-1 helps tumor cells evade the immune system - COX inhibitors like aspirin...therapeutics such as  (Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in ...combination of the CRISPR/Cas9 methodology with T cell receptor engineering (Ref 5)  as described by Blue...

  16. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...reagents and keep the cells happy. Harvesting virus Once you have transfected your cells, wait 2–5 days and...however, you’ve collected the producer cells, you’ll need to lyse the cells to let the virions out. We recommend...particles that can transduce cells, which can vary based on target cells and can be altered by freeze/...come into contact with the rAAV and apply common cell culture practices diligently throughout the production...capsid genes.  In this helper-free system, HEK293T cells, expressing Ad E1A and E1B, can be transfected with...and then collect the supernatant and/or producer cells for further processing. What component(s) you need...released into the culture media, while others remain cell-associated (Vandenberghe et al., 2010). For certain...

  17. Hot Plasmids - November 2023

    Type
    Blog Post
    ... between living cells by TransitID. Cell, 186(15), 3307–3324.e30. doi: 10.1016/j.cell.2023.05.044. PMID...lentivirus and MMLV gag, to specific cell types. It can also make CAR-T cell therapy more accessible by simplifying... efficient prime editors. Cell, 186(18), 3983–4002.e26. doi: 10.1016/j.cell.2023.07.039. PMID: 37657419...called DIRECTED (Delivery to Intended REcipient Cells Through Envelope Design). This modular approach ...viral surface, expanding the range of targetable cell types. DIRECTED is compatible with fusogens from...process and could even enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can ...DIRECTED plasmids here! Strebinger, D., et al. (2023). Cell type-specific delivery by modular envelope design...

  18. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at...either in whole cells or as a pool of fusion proteins localized to a specific region of the cell. For example...the relationship between the age of a cell, protein, or cellular structure and a particular biological...trafficking to a subcellular location, start of gene expression, development of a cell structure, etc)....timers, researchers can look at a single image of a cell and understand how protein localization changes ...was a tetramer with a propensity to aggregate in cells (Tsuboi et al.), limiting its use. Nonetheless, ...versatile timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus embryos. Of course...

  19. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...packaging AAV is transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids...precipitation. Instead of sonicating the cell pellet, I lyse the cells with this AAV lysis buffer and freeze...bath to release the virus particles from the cells. Cell lysates are then treated with benzonase immediately...five days after transfection, the AAV-containing cells and media are harvested and purified. In total, ...not counting the time needed to expand the HEK293 cells. Check out the Addgene AAV Production protocol for... strains, like NEB Stable. NEB Stable competent cells lack RecA, a protein that aids in the homologous... AAV. I skip the PEG precipitation of the HEK293 cell culture supernatant because it’s time consuming ...

  20. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ... for stem cells, or metastasized tumor cells, or alzheimers-affected neurons, or [your cell line of interest...critical to stem cell survival. Knowing which genes influence the survival of our cell culture is the foundation...minimal side-effects on healthy cells and maximal effect on unhealthy cells. Until recently, the human genome...ENCODE project, find enhancers in embryonic stem cells, and conclude with a targeted CRISPR screen that...essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds just over 13,000 significant...We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus...The result is a heterogenous population of stem cells, usually housed in a single flask. Through a bit...
Showing: 181 - 200 of 813 results