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We narrowed to 813 results for: cell

Showing: 281 - 300 of 813 results

  1. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor and sometimes ...transduction efficiency, and is an excellent choice for targeting cardiac cells (Bish et al., 2008). It can ... Therapy: An Overview. Cells, 12(5), 785. https://doi.org/10.3390/cells12050785 Kaludov, N., Brown, K....delivery across species. Cell, 184(19), 4919-4938.e22. https://doi.org/10.1016/j.cell.2021.08.028 Tervo, D...tissue tropism, allowing them to target particular cell types. Figure 1: Diagram of AAV components...key that lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic...multiple coreceptors, which may be present on different cell types and sometimes vary between target species ...

  2. NETRF

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    Collection
    ... Modeling Neuroendocrine Tumors Using Adult Stem Cell-Derived Organoids Matthew Meyerson Dana-Farber Cancer...Institute Epigenetic Regulators of Intestinal Endocrine Cells and Carcinoid Tumors Qiao Zhou Harvard University...University Epigenetic Regulators of Intestinal Endocrine Cells and Carcinoid Tumors Bradley Bernstein Broad Institute...

  3. What's New in CRISPR - November 2021

    Type
    Blog Post
    ... into AAV particles for delivery into mammalian cells, especially when fused to effector proteins for ..., base editing, and genome editing in mammalian cells. The “V4” version (named CasMINI) had the highest...an active nuclease.  Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR editing...effective in genome editing in both bacteria and human cells using various delivery methods, including plasmid...mRNA or Cas9 mRNA and sgRNA, in human and mouse cells. They further develop a version of SEND for gene...modular system for delivering gene editing tools into cells. Image adapted from Segel et al. 2021.   Prime...correct mutations in a variety of target genes and cell types. Their Nature Biotechnology paper showcases...

  4. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...of Multiple Kinase Activities in Live Single Cells." Cell 157, no. 7 (2014): 1724-734. PubMed PMID: 24949979...Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to ... dimeric Dronpa is less prone to aggregation in cells compared to the tetrameric Dronpa it was derived...inhibitors, psMEK1 and ERK KTR-mRuby2 expressing cells were incubated with an inhibitor of choice, exposed...inactivation, and then decreases. Treatment of the cells with a PP1/PP2A inhibitor leads to maintenance of...Raf-MEK-ERK signaling in the hypodermis and gut cells of C. elegans causes the same phenotype. Activation...signaling pathway with psRaf1 or psMEK1 in these same cells by 24-48 hours exposure to blue light also induced...

  5. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...intensity evolves over time in living cells at a power used for live-cell imaging. To conclude, the most photostable...molecule will photobleach at some point. For live-cell imaging, it is desirable to have fluorescent proteins...experiment. The best fluorescent proteins for live cell imaging can be excited many times, thereby producing...protein is a key requirement for quantitative live-cell imaging, it is important to be able to quantify ...it is advisable to perform time-lapse imaging of cells producing the fluorescent protein of interest under...natural situation. Measuring photostability in living cells provides a much more realistic view of photostability...spectral class on the microscopy system and in the cell type (or tissue) that is intended to be used with...

  6. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ... protein (CcdA) that protects the cell against the toxic CcdB. Cells that lose ccdA through the loss of...disrupted allowing cells expressing the recombinant plasmid to propagate. Any cells that contained non-recombinant...which is responsible for plasmid maintenance during cell division. ccdB codes for the toxic protein (CcdB...broken double stranded DNA and ultimately causing cell death. ccdA, another gene found in the ccd operon...Gateway takes advantage of the same principle that cells will not propagate while expressing the gene. Briefly...can one work with a plasmid/gene that kills the cells expressing it? The answer lies in special strains...PMID: 8254658. 5. Bernard, P. and Couturier, M. “Cell killing by the F plasmid CcdB protein involves poisoning...

  7. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ...Targeted Human Stem Cell Clones Detected by Whole-Genome Sequencing. Cell Stem Cell 15:27–30 . https://...protein, in order to mitigate the ability of the cell to use an alternative ATG downstream of the annotated...process, and usually involves the need for single cell cloning and subsequent screening for successful ...standard requires not one but two rounds of single cell cloning – as a control, one should revert the edit...passenger variant that came along with the single cell clone (although this is rarely done). When targeting...levels of modification. Whole-genome sequencing of cells modified by CRISPR indicates that the consequences...mutations (Veres et al., 2014). When working with single-cell clones, the authors note that “clonal heterogeneity...

  8. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...Purification Process with pCri Cell-free protein expression kit Cell-free expression systems have been...testing and comparison of different cell-free expression systems; the Cell-free expression test kit represents...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of...: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq PUF Assembly...facilitate gene expression in bacteria, yeast, mammalian cells, insects, and even Xenopus oocytes. The expression...of proteins in both prokaryotic and eukaryotic cell extracts (Gagoski et al 2015). He has also constructed...to perform statistically relevant benchmarking of cell-free expression systems, and tested for product ...

  9. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...percentage of total live cells are GFP+. In this plot, with adjusted voltage, cells that did not receive ...would serve you well. Percentage of cells in a parent gate (all live cells for example) that are positive ...example, you have introduced a GFP protein into your cells (visualized via the FITC channel) and you are trying...graphs could simply be the percentage of the total cells present in the gate.   Figure 1: Example...looking at proteins that are expressed regularly in cells and might be upregulated or downregulated based ...populations. MFI is the average brightness from all cells that are positive for the marker of interest. MFI...

  10. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    ... to express one gene in one cell type and another gene in a different cell type (6–8). Although it’s often...systematically study gene function with temporal control and cell-type specificity! The Gal4/UAS system At a more...the hedgehog gene can be overexpressed in several cell types by using different Gal4 drivers. It is much...much easier to mate flies to get the right cell type expression as opposed to making a new fly with hedgehog... Targeted gene expression as a means of altering cell fates and generating dominant phenotypes., Development...Neuro-Trauma during Asymptomatic Viral Infections, Cell Host Microbe 22, 48-60 (2017). PubMed PMID: 28704652...expression, lineage tracing, and mosaic analysis., Cell 141, 536–48 (2010). PubMed PMID: 20434990. PubMed...

  11. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...replicating too quickly can slow cell growth or even kill the host cell. For stringently controlled plasmids...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...be synchronized with the replication of the host cell's chromosomal DNA.  Plasmids are said to be under... stringent control when they depend on the host cell’s initiation proteins to start their own replication... occurs at the appropriate time—for example, the cell must have enough nutrients available to complete...the amount of active DnaA available is one of a cell’s best tools to regulate DNA replication. DnaA negatively...energy source, ATP binding is one more way for the cell to control DNA replication. A variety of other proteins...

  12. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...splitting my cells first. Time management through planning I probably wouldn’t have fallen into my cell splitting...for Scientists. Back when I used to do mammalian cell culture, I would often respond to friends asking...there in a few minutes, I just need to split some cells.” Inevitably, this task would take longer than I...since my days as an undergrad working with cancer cell lines. As I started planning out my own experiments.... In my anecdote above, I prioritized splitting cells over meeting up with friends, but didn’t realize...mean in terms of the utility of your tasks. In my cell splitting anecdote, I probably could have grabbed...with friends and come back to the lab to split my cells later. Eating with friends has a somewhat immediate...

  13. Depositor Collections

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    Collection
    ...Collection Allen Institute for Cell Science Plasmid Collection New England BioLabs Cell-Imaging Plasmid Collection...Botman-Teusink Yeast FP Collection Protein Expression Cell Migration Consortium Malate Dehydrogenase CUREs ...

  14. AAVs in Retinal Gene Therapy

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    Blog Post
    ...expression in non-dividing cells because they remain in episomal form in the cell’s nucleus. But since nothing...hope are many more clinical successes for gene (and cell) therapies. One may wonder why the first AAV-based... providing long-term gene expression in retinal cells - AAV - were becoming available. At the same time... lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of choice for ocular...long term expression in dividing and non-dividing cells due to its (random) integration into the genome,...limited tropism in the retina, primarily infecting cells of the retinal pigmented epithelium (RPE). AAVs... for patients, this gene can be targeted to RPE cells using a lentiviral vector and a clinical trial is...

  15. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...when trying to screen a limited number of cells like primary cells or in vivo. Recently, the labs of David... cultured cells as well as more difficult conditions such as model systems and primary cells. References...with both A375 (melanoma) and AT29 (colon cancer) cells and assessed the libraries performance via the depletion...important criterion for probing a limited number of cells. Get the Brunello CRISPRko sgRNA library as pooled...subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition (CRISPRi) libraries...study screening for vemurafenib resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation...CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647-661. PubMed PMID: 25307932. PubMed...

  16. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially if you...reporters allow you to easily screen or select for cells that are expressing your gene of interest at high...promoter, multicistronic plasmids ensure that any cells that are positive for your marker should also be...anytime you want to express multiple genes in the same cell. Although it is possible to drive co-expression ...promotes translation of the viral genome in eukaryotic cells.1,2 Since then, a variety of IRES sequences have... cistron due to factors such as the experimental cell type and the specific genes cloned into the vector...scientists. 2A cleavage is universal in eukaryotic cells, and, although some scientists report close to 100%...

  17. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    ...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed ...Cepko Interested in Working on the Eye 9:05-10:16 : Cell Targeting in Retinal Gene Therapies 10:17-14:34 ...28:39 : Methods for Delivering Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools...publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation with GFP-dependent Cre...

  18. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ...co-expression among cellular markers You’ll also need to know if markers are co-expressed in your cells when designing...immunologists, allowing for the rapid analysis of cell populations within a heterogenous tissue type, such...methods, the identification and isolation of rare cell types. In recent years, full spectrum flow cytometry...Spectral flow allows for autofluorescence extraction Cells produce a low level of fluorescence called autofluorescence...single color control. Reference controls: Beads or cells? Compensation beads are a convenient tool to obtain...you prepare reference controls of both beads and cells for a single fluorophore. Although compensation ...Additionally, it is critical to include unstained beads or cells within your reference group to ensure proper unmixing...

  19. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    ... Ba/F3 and MCF10A cellular models. Ba/F3 and MCF10A are growth factor dependent cell lines and any mutations...attempting to characterize mutations in a pooled cell population, as mutations with strong phenotypic ...functional groups, based on how each one impacted cell viability (see figure). Most of the mutations were...to be “neutral,” having no significant effect on cell viability compared to a wild-type control. Approximately... the mutations, however, significantly increased cell viability, and these “activators” may be the direct...annotation of somatic mutations in cancer." Cancer Cell 33.3 (2018): 450-462. PubMed PMID: 29533785. PubMed...

  20. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...fluorescence images of C2C12 cells stably expressing pTRE-Tight-MitoTimer and rtTA. The cells were pulsed with 2µg... Robinson et al., Dev Cell 2010 Feb 16;18(2):324-31. Cheeseman et al., J Cell Sci 2013 May 1;126(Pt ...neighboring cells in imaging applications and the mapping of neuronal projections to their associated cell bodies... targeted to subcellular regions or protein complexes for proteomic mapping in live cells. Lam et al...been used by several groups to monitor protein and cell ageing, as the spectrum shift occurs during Timer...(pTRE-Tight-MitoTimer) to show its usefulness in cell culture to report changes in mitochondrial turnover...rapamycin-binding domains, such as FKBP and FRB. The cell does not appear to mind extraneous proteins bound...
Showing: 281 - 300 of 813 results