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We narrowed to 859 results for: cell

Showing: 481 - 500 of 859 results
  1. Understanding GPCR Signaling Using TRUPATH

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    Blog Post
    ... to environmental conditions, cells must be able to detect extracellular stimuli. One way they do so is...complex converts a signal received by a GPCR on the cell’s surface into action that can result in the initiation...protein transducers. To use the TRUPATH system, cells are transfected with plasmids expressing the GPCR... and the appropriate Gγ-GFP2 and Gβ constructs. Cells are then treated with the luminescent enzyme substrate...bound to the intracellular surface of the GPCR. Binding of a ligand to the extracellular surface of the...number of signaling cascades, inducing different cellular responses. One of the biggest advantages of such...
  2. Antibodies 101: Epitope Availability

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    Blog Post
    ... have been extracted from a cell, instead of when it is attached to a cell membrane.  Picking the Right...epitopes that are available on the outside of the cell. The same protein may have different epitopes available...need to validate on denatured proteins or on live cells. But for others, you may need to validate that it...
  3. Fluorescent Biosensors for Measuring Autophagic Flux

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    Blog Post
    ...the vacuole in yeast or the lysosome in mammalian cells. This decrease in pH quenches SEP but not DsRed,...acidity, this biosensor can’t be used with fixed cells. Find the mitochondria-targeted Keima variant at...Levine. "Methods in mammalian autophagy research." Cell 140.3 (2010): 313-326. PubMed PMID: 20144757. PubMedCentral...measurements of macroautophagy and mitophagy in mammalian cells using a single fluorescent biosensor." Scientific...
  4. 10 Great Guest Posts We're Thankful For

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    Blog Post
    ...Your Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts...which specific chemical inputs are received by a cell. These so-called recombinase-based state machines...used to deliver a genes of interest to a variety of cell types in-vivo. These are widely used by the optogenetics...activation and repression of different genes in a single cell, and localization of a fluorescent signal to your... Below you’ll find brief summaries of several excellent guest posts from our depositors. Interested in...
  5. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

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    Blog Post
    ...Nanobody toolkit to monitor retrograde transport at the cell surface Article contributed by Marcy Patrick  Listen...nanobodies to study retrograde transport from the cell surface. Their toolkit includes a set of plasmids...new technologies to quantify proteins in tissues, cells, and biofluids. Recently, Sam Myers, a Scientist...an sgRNA expressed from a separate plasmid. When cells are treated with biotin–phenol and H2O2, APEX2 labels...
  6. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

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    Blog Post
    ... be stable in the cell. gDNA guides will also need to be transfected into the cells of interest. In contrast...unable to show TtAgo-mediated cleavage in mammalian cells, likely due to high temperature requirements. To...plasmid and genomic DNA in a variety of mammalian cell lines, with an efficiency similar to that of Cas9...
  7. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

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    Blog Post
    ...R-factors could be transferred between bacteria via cell-to-cell contact; however, scientists noted that, unlike...cytoplasmic factors that could be transferred between cells. At the time, these extranuclear agents of heredity...PMCID: PMC3785787. References: Lederberg, Joshua. "Cell Genetics and Hereditary Symbiosis" Physiological...
  8. Water Bath Protocol

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    Protocol
    ...restriction digests needed in molecular biology, and thaw cell lines. There are many different types of water baths...
  9. Water Bath Protocol

    Type
    Protocol
    ...restriction digests needed in molecular biology, and thaw cell lines. There are many different types of water baths...
  10. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...By taking advantage of repair pathways within the cell — in some cases, the same repair pathways that were...adenosine (A) to inosine (I, treated as G by the cell). Surprisingly, the team noticed that this editor...cytosine editing between bacteria and mammalian cells Researchers in the Xueli Zhang lab were making similar...2021). Their GBE produced C → G edits in HEK293T cells with over 50% efficiency in 23 out of 30 target ...follow the same initial mechanism as in mammalian cells — by excising the freshly edited U bases — but E...these fusion constructs improved editing in various cell types, although efficacy varied among target sequences...CGBEs at different target sequences and in different cell types, available at www.crisprbehive.design. CGBEs...
  11. A Primer on Optogenetics: Introduction and Opsin Delivery

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    Blog Post
    ...opsin expression in the intended brain region and/or cell type (I’ll talk more about this in detail later)...desired optogenetic construct in the brain region and cell type of interest is obviously of critical importance...wish to stimulate or inhibit neuronal firing, the cell-type specificity, the kinetics of the opsin variant...control of a generic promoter). A DIO opsin allows cell type specificity dependent on the promoter of the...expression of your opsin is no longer required to achieve cell-type specific opsin expression. However, the types...spread, and with co-localization immunofluorescence, cell-type specificity. Viruses that only express the ...experimentation.  Whether you attempt to stimulate the cell body or an axon terminal also affects the length...
  12. Generating Mouse Models Using CRISPR/Cas9

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    Blog Post
    ... into a mouse embryonic stem cell (ESC) line, enrich and select for cells that had successfully incorporated... when it gets exciting – the cell sends out an “SOS” signal and cellular repair mechanisms rush in to ...Cas-Mediated Genome Engineering. Cell 154:1370–1379 . https://doi.org/10.1016/j.cell.2013.08.022 Yoon Y, Wang ...Cas-Mediated Genome Engineering. Cell 153:910–918 . https://doi.org/10.1016/j.cell.2013.04.025 Zhang X, Liang...develop into chimeric mice (with a fraction of the cells in the adult mice derived from the engineered ESCs...Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR/Zygote Injection Approaches...
  13. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ... Guide RNA and Target DNA. Cell 156:935–949. https://doi.org/10.1016/j.cell.2014.02.001  Ran FA, Cong ... Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell.2020.03.023 Additional Resources...acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency ...package each piece into an individual AAV. When a cell is transduced by both AAVs, the full length gene...SpCas9 which had modest editing rates in HEK-293T cells when compared to the full-length SpCas9. In 2016... allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: ....
  14. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...interest live within a cell,” “Does this disease change the architecture of my cells,” or “How does this ...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...IF), is an immunoassay that brings to light the cellular world. The technique allows you to ask questions...
  15. CRISPR-based Genome Editing Systems For Editing in E. coli

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    Blog Post
    ...construct and then introduce both plasmids into the cell. The edit in the chromosome can now take place. .... After editing, pTargetF can be cured from the cell by inducing expression of the sgRNA that targets ...all the edits you want, cure pCas9 by growing the cells at 37 °C.     The new pEcCas/pEcgRNA system We ... to take place. Then plasmids are cured from the cell. Steps 1-1 and 1-2 can be done simultaneously. Image...tools for faster construction or reprogramming of cells to understand the genetic and biochemical mechanisms...phenotypes and to design a new generation of engineered cells for green chemistry and biomedical applications.... Chinese Academy of Sciences (CAS) Center for Excellence in Molecular Plant Sciences in 2019. She joined...
  16. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ... want to make sure your cells have first reached an appropriate density. Cells in the exponential growth...collecting dead cells, and, conversely, you cannot induce too early as there are not enough cells in the culture...host protein, using up valuable resources in the cell thus leading to slowed growth. Additionally, some...such as TB or 2XYT is optimal because of the high cell-density they support; however, minimal media supplemented...
  17. Keeping Up With CRISPR/Cas9

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    Blog Post
    ...RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014 January 30. (Pubmed) Programmable...correct disease mutations, both in mice and in human cells. But that important advance was quickly followed...
  18. Plasmids 101: Gram Negative and Positive Bacteria

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    Blog Post
    ...negative bacteria is due to differences in cell wall composition. Cell wall structure Gram positive bacteria...complex within the peptidoglycan layer, staining the cell purple. When the alcohol is added, it dehydrates...Silhavy, TJ., Kahne, D. & Walker S. The Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414...
  19. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

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    Blog Post
    ...editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652.e29. https...efficient prime editors. Cell, 186(18), 3983-4002.e26. https://doi.org/10.1016/j.cell.2023.07.039 Lin, Q.,...correct only a minority of those variants in most cell types. But prime editing, a CRISPR technique developed...unedited DNA. The heteroduplex is resolved by the cell’s mismatch repair system, and more recent prime editing...editing demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations...type of edit, genomic context, delivery strategy, cell type, and goal of the experiment can all impose ...nicking the non-edited strand, they can cause the cell to remake that strand using the edited strand as...
  20. What's New in CRISPR - May 2019

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    Blog Post
    ...genome-wide deamination of RNA cytosines in human cells. Thus, they engineered two CBE variants with mutations...RNA edits by >390 fold and >3,800 fold in human cells with no significant reduction in editing efficiency...mice and in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR...
Showing: 481 - 500 of 859 results