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We narrowed to 876 results for: cel

Showing: 501 - 520 of 876 results

  1. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...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...

  2. Scientists Map the SARS-CoV-2-Human Interaction Network

    Type
    Blog Post
    ... 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...

  3. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ... 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...

  4. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...to quantify rates of mutation in a bulk or single cell-derived genomic DNA sample as part of a precision... successfully analyzed gDNA from a few thousand cells from single wells of a 96-well plate without prior... that they are easily able to distinguish single-cell derived samples with mono-allelic mutations versus...Isolation of single-base genome-edited human iPS cells without antibiotic selection. Nat Meth 11, 291–293... and Highly Specific Screening of Genome Edited Cells. PLoS ONE 11, e0153901–17 (2016). PubMed PMID: 27089539...mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology 31, 822–826 (2013). PubMed...

  5. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ...genetic cargo, typically an expression construct, to cells/organisms. This is often cDNA, shRNA, or something...perform very similar functions to infiltrate the host cell and deposit viral cargo. Gene expression While AAV...what will be delivered and expressed in the host cell from the transfer plasmid. Viral packaging and entry...altered tropism of AAV to target different tissue or cell types. AAV also requires some back up from adenovirus...promoter that you wish to deliver to your target cells. In adenovirus, the ITRs serve as a type of origin...deficient. This is typically supplied by the production cell line (HEK-293) during production. Fun fact: There...

  6. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...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...

  7. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    ...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...

  8. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    ...proteins that bind an RNA motif of interest in living cells. Challenges with existing RNA-protein interaction...and naturally occurring complexes present in the cell, it can also introduce non-physiological RNA-protein...a short timescale, such as during drug action or cell signaling. Limitations of RaPID Like other BirA*... controls include both scrambled RNA motifs and cells transfected with BirA* protein alone (i.e. no RNA...protein that’s highly expressed in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels...2018). RNA–protein interaction detection in living cells. Nature Methods, 15, 207-212. PubMed PMID: 29400715...

  9. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ...ssODN repair template in induced pluripotent stem cells made from the patient. With the first gRNA, they...homology-directed repair (HDR) taking place in 41.2% of edited cells. The results were even lower for the second gRNA...13.1% targeting efficiency with only 13% of edited cells employing HDR. Ma et al. moved forward with gRNA...as whole exome sequencing of embryo derived-ES cells.They also conducted whole genome sequencing in selected...babies.” The differences in editing between iPS cells and embryos make a case for conducting research ...analysis is not possible with IVF, as most of the cells are needed for proper development. Ma et al.’s experiments...

  10. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ... Mammalian Autophagy Research. Cell,140(3), 313-326. doi:10.1016/j.cell.2010.01.028. PubMed PMID: 20144757...analyzing autophagy of cytosol and organelles in yeast cells. Autophagy (Greek for “self-eating”) is induced ...radio-labeled long-lived proteins as a proxy for autophagy. Cells are cultured with isotope-labeled amino acids for...fluorescence regardless of its localization in the cell. SEP’s pH sensitivity means Rosella will fluoresce...measurements of macroautophagy and mitophagy in mammalian cells using a single fluorescent biosensor. Scientific...

  11. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...transgenic animals, embryonic stem cells, and/or tissue-specific cell types: Cre-lox recombination. What...downstream of promoters that are active only in certain cell or tissue types, during certain stages of development... recombinase can be expressed only in specified cells or at specified times. Combining this with some ...has been used to successfully introduce Cre into cells of interest. What if I need two separate recombination...2. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Sauer...

  12. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...specific cell types or entire organs in planarians. For example, the gene Piwi labels neoblast cells and the...techniques to alter gene expression or to label specific cells, but there are too few resources to teach students... in tissue. This is useful for labeling specific cell types and for viewing how gene expression changes...

  13. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable for up to...possible and submitted our manuscript to the journal Cell just six weeks later. The Zika publication timeline...of Toronto. His research is focused on developing cell-free synthetic biology applications for human health...Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from...de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939. PubMed PMID: 25417166. PubMed...Keith, et al. "Paper-based synthetic gene networks." Cell 159.4 (2014): 940-954. PubMed PMID: 25417167. PubMed...

  14. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...purpose and mechanism of polyadenylation vary across cell types, but polyadenylation generally serves to promote... thought that poly(A)s are used to control the cellular concentration of regulatory RNAs and may additionally...additionally act as a quality control mechanism to rid the cell of mis-folded RNAs. Eukaryotic termination and polyadenylation...signal by using in vitro analyses. Molecular and Cellular Biology. 1992. PMID: 1333042. PubMed Central PMCID...efficient rabbit beta-globin mRNA 3' end formation. Cell. 1987. PMID: 3568131.  Hager S, Frame FM, Collins...

  15. Selecting Your Plasmid Purification Kit

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    Blog Post
    ... For most kits, the steps for lysing cells and removing cellular debris look very similar. However, keep...relative term — even armed with the highest-competency cells you will still struggle to get transformants if ...quantity) – Depending on the experiment, mammalian cell transfections can require quite large amounts of...your plasmid; separate plasmid DNA from unwanted cellular “debris”; and then clean-up the plasmid DNA (Figure...your bacteria to release the plasmid, separate the cell debris from the plasmid, and then clean-up the plasmid...Kits can be optimized based on the plasmid size, cell model, or overall throughput needed. Large constructs...

  16. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Single-Domain Camelid Antibodies. Cell 181:1004–1015.e15 . https://doi.org/10.1016/j.cell.2020.04.031 Zimmermann ...coronaviruses and enable the virus to enter host cells. To do this, they injected the spike protein from...camelid antibodies that bind to the spike proteins in cell culture, and inhibit these viruses.  But llama ...hidden in the molecular crevices proteins on the cell surface. Figure 1: Comparison of common antibody...blog Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification RANbodies: Reporter...

  17. Of Myc and Men

    Type
    Blog Post
    ...For about as long as the disciplines have existed, cell and molecular biologists have been continuously ... peer into the mysterious, microscopic world of cells. The development of antibodies as tools allowed ...monoclonals would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different...specific for human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616. https://doi.org/10.1128/mcb.5.12.3610... and immunoglobulin heavy chain binding protein. Cell 46:291–300. https://doi.org/10.1016/0092-8674(86...signal prevents secretion of luminal ER proteins. Cell 48:899–907. https://doi.org/10.1016/0092-8674(87...

  18. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ...What works well in one cell or neuron type may not work well for a neighboring cell type, even within the...options while AAV8 (82%) and AAV9 (84%) were both excellent at infecting our target neurons (Figure 1). However... conditions (3,5,8), injection titers (1,2), and cell types used for packaging viral particles (6), can... vector performance in various brain regions and cell types. Power in numbers A Pubmed search for “AAV...and Share AAV Data with Addgene's New AAV Data HubCelebrate Open Data Day with Addgene’s AAV Data Hub!Data...

  19. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ... reader and an Excel spreadsheet. Start by scanning the sample barcodes into the cells in the same order...using an electric charge to help the DNA enter the cells. Some samples require a reduced antibiotic concentration...concentration or need to be transformed into different cells than were originally requested. Most of our samples...tubes and use this ID to name the corresponding Excel sheet.3. Be specific  - When assigning a physical...storage process - from deposited DNA to transformed cells and finally to bacterial glycerol stocks that are...

  20. 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...
Showing: 501 - 520 of 876 results