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We narrowed to 819 results for: cell

Showing: 181 - 200 of 819 results

  1. 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...

  2. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ... a big issue for cells, but it may confound research results or even kill your cells if your gene of interest...necessary to replicate plasmids and make sure the cells maintains it. Here, we'll cover the promoter, which... Because transcription machinery differs between cell types or organisms, promoters must be similarly ...variable. Bacterial promoters only work in prokaryotic cells and typically only in the same or closely related... were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc) require unique...complex, having fewer parts than those in eukaryotic cells. Some promoters are constitutively active and on... at will with a chemical, heat, or light. In the cell, promoters themselves are controlled by still other...

  3. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

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    Blog Post
    ...produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells are fused with activated...activated B cells from the spleen of mice injected with the antigen of interest. The immortalized cells, called...distributing over 3,000 antibodies and associated hybridoma cell lines. The DSHB was created by the National Institute... individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing...

  4. Viral Vector Guides and Plasmids

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    ...A) Wild-type viruses infect cells in an organism and use the host cellular machinery to produce more virions...they can be delivered to packaging cell lines and use their cellular machinery to generate viral vectors...In this way, viruses are excellent vehicles to deliver genetic cargo into cells. To create safe and effective...popular tools for delivering genetic information to cells. These vectors rely on production plasmids that ... ability to transduce dividing and non-dividing cells. These virusal vectors enable long-term stable gene...transduction efficiency and only infect dividing cells. They integrate permanently into the host genome...Retroviral vectors are popular for making stable cell lines or clinical use. Adenoviral Vector Guide |...

  5. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

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    Blog Post
    ...help in the capture of other rare cell types, like stem cells or cells infected with malaria or other parasites...tumor markers to subsequently capture cancerous cells expressing low levels of tumor antigen. I asked ...looking at, magnetic isolation of circulating tumor cells, we found that the binding stability really mattered... in the pili that they use to anchor on to host cells. Addgene: Were there any major hurdles along the... donor for keeping going while we worked out the cell capture system.) Cancer and other applications Addgene...cancer? Howarth: The capture of circulating tumor cells (CTCs) is going to have a big impact on our understanding...polymerizes affibodies to enhance magnetic cancer cell capture. Fierer et al (Proc Natl Acad Sci U S A....

  6. 27 Hot Plasmids from 2016

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    Blog Post
    ... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ...plasmid to generate a population of mutant cells. The mutant cells were treated with WNV at a dose and duration...regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3, EMC2, DERL2, UBE2G2...AID-tagging system to essential genes in human cells and mouse ES cells because it was difficult to fuse the AID...-to-study genes in hard-to-study cell types. Natsume T, et al. Cell Rep. 2016. PubMed PMID: 27052166 ...small GTPases involved in cell signaling pathways that control many cellular processes such as differentiation...therefore varies with cellular location: it is both red and green through much of the cell, but turns more ...

  7. Fluorescent Protein Guide: Biosensors

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    ... pH Imaging intracellular pH in live cells (pHRed) Imaging Intracellular pH in Live Cells with a Genetically...SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation. Cell Rep. 2019 Jun 25;27(13):...biosensor for visualization of acetyl-CoA in live cells. Cell Chem Biol. 2025 Jan 21: S2451-9456(25)00002-9... Tantama ATP QUEEN ATP sensors for live-cell imaging Live cell imaging of metabolic heterogeneity by quantitative...for Imaging Insulin Secretion in Single beta Cells. Cell Chem Biol. 2017 Apr 20;24(4):525-531.e4. Dmytro...accessibility. Cell. 2022 Sep 1;185(18):3390-3407.e18. David Clapham Return to top Cell Physiology Target...ATP (extracellular) ecAT3.10 extracellular ATP sensor (cell surface targeted) Imaging extracellular ATP...

  8. Five Popular Model Organisms

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    Blog Post
    ... properties and processes with our own cells. Like human cells, yeast DNA is packaged into chromosomes...differentiation of β-cells in the the developing zebrafish pancreas. They first tagged these cells using plasmids...recapitulate the complex interactions between cancer cells, therapeutic drugs, and the rest of the body than...than studies in other model organisms or cell culture. For example, Scott Lowe’s lab utilized mice to study...to identify a set of mutations that made cancer cells more resistant to chemotherapy. You can find these...to optimally control gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces...is very amenable to genetic manipulation. Yeast cells are great model organism not only for the reasons...

  9. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

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    Blog Post
    ...manipulations on cells, and has potential use for a wide range of applications in cellular biology. Van ... of a microtubule-associated protein to regulate cell growth dynamics Article contributed by Klaus Wanisch...of the modified EB1 protein are overexpressed in cells, microtubule growth can be switched off with blue...paper demonstrated that the migration of a cancer cell line equipped with photodissociable EB1 could be...Van Haren J et al. Nature Cell Biology 2018 Pubmed PMID 29379139. The “Riboglow” platform for visualizing...used to visualize RNA localization in mammalian cells. These tools consist of cobalamin-binding riboswitches...proteins (FP) with improved tissue penetrance and live cell imaging capabilities. Upon developing a new imaging-based...

  10. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with cell-based method ...Requires large cell population, senstive to time since cell fixing HTGTS Not reported Cell-based Identifies...method IDLV 1% Cell-based Programmable, can detect DSBs in live cells Not as senstive as other unbiased methods... use single cell clones for in vitro CRISPR experiments. The likelihood that a single cell clone derived...picture of the impact of CRISPR editing in the model cell line or organism. Several options exist for unbiased...Osborn et al. 2016) and HTGTS (Frock et al. 2015) for cell-based experiments. These strategies can be used ...off-target editing events. Two of the most common cell-based methods are genome-wide, unbiased identification...

  11. All in a Twist: dsRNA

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    Blog Post
    ...mammalian cells. Here, we will cover how these structures are generated, why it’s important for cells to sense...., Hur, S. Cellular origins of dsRNA, their recognition and consequences. Nat Rev Mol Cell Biol, 23: 286...performs almost all of its biological functions in our cells in the single strand form, but double stranded RNA...with bidirectional transcription. In an uninfected cell, dsRNA rarely circulates, but upon viral infection...Unlike dsRNA, DNA:RNA hybrids are abundant in normal cells and are arise from a ssRNA engaging one strand of...genes is constantly occurring throughout a single cell, so these nucleic acid hybrids are always around...at risk. Consequences of duplexed RNA Mammalian cells have multiple receptors in place to sense dsRNA ...

  12. Antibodies 101: Designing Your First Flow Panel

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    Blog Post
    ...and inside the cells and are thus typically used for distinguishing live from dead cells. The combination...say I am culturing HEK 293T cells in vitro. I transduced the cultured cells with a virus that encodes a...panel. Besides some background cell death constantly happening in cell culture, the transduction process... to some cell death. As a result, there is a high abundance of positively stained dead cells present and...When analyzing your cells using flow cytometry, you are typically measuring the presence or absence of...certain markers on the surface or the inside of your cells. While proteins themselves can emit intrinsic fluorescence...Another way to label proteins is through treating cells with fluorophore-conjugated antibodies that attach...

  13. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ...transfected into a packaging cell line, usually a derivative of 293T. Packaging cells are incubated and after...fine-tuning selection and expression in your target cell line, researchers often find themselves faced with...impact the transfection efficiency of the packaging cell line and thereby affect the titer of the virus. ...layer flask will require some initial optimization; cell number, media volume, and plasmid concentrations...larger vessel to the 10 cm2 dish and adjusting the cell number, plasmid concentration, and experimental ... consideration when titering a lentivirus is the cell line that is used for titering. Typically researchers...researchers will choose a relatively permissive cell line for titering in order to obtain the best titer possible...

  14. Viral Vectors 101: Systemic Capsids

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    ... your cell type of interest, while miRNA target sites can be used to prevent expression in cell types ...endothelial cells in rodent CNS AAV-PHP.V1 shows higher transduction in CNS endothelial cells (60%) and ...endothelial cells in the retina and spinal cord (Krolak et al., 2022). Systemic capsids for broad cell type ...delivery across species. Cell, 184(19), 4919-4938.e22. https://doi.org/10.1016/j.cell.2021.08.028 Vormstein-Schneider...capsid, (b) cargo, and (c) delivery method determine cell type- or region-specific transgene expression. (...include variants with broad CNS or PNS transduction, cell type specificity, or retrograde transport in neurons...sequences, denoted by triangles] facilitate tissue or cell type specificity. An on/off genetic switch can be...

  15. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...thought to activate programmed cell death or a dormant state for bacterial cells that have been infected with...bacterial cells, LwaCas13a does not display nonspecific RNA cleavage activity in eukaryotic cells. They arrived...target RNAs, although its usefulness in bacterial cells will likely be limited by the propensity of Cas13a...pool of RNAs) or studying RNA processing in live cells. Indeed, groups have utilized dCas13 fusion proteins...genotype human DNA, identify tumor mutations within cell-free genomic DNA, and detect COVID-19 (Joung, Ladha...Ladha et al., 2020). Cas13a in mammalian cells The Zhang lab made the exciting discovery that some Cas13a...Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They characterized Cas13a...

  16. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...find both single-allelic and bi-allelic cells. Single-allelic cells usually make up the majority in culture...allows you to sort for GFP-positive cells and to enrich for those cells that were positively transfected....pluripotent stem cells differing exclusively at two early onset Parkinson Point mutations. Cell, 146(2), 318...Double-Strand Break Repair in Mammalian Cells. Molecular and Cellular Biology, 18(1), 93–101. https://doi....genome editing specificity. Cell, 154(6), 1380–1389. https://doi.org/10.1016/j.cell.2013.08.021  Ran, F. ...CAS-Mediated Genome Engineering. Cell, 153(4), 910–918. https://doi.org/10.1016/j.cell.2013.04.025  Yang, H....CAS-Mediated Genome Engineering. Cell, 154(6), 1370–1379. https://doi.org/10.1016/j.cell.2013.08.022  Additional...

  17. Viral Production

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    ... preparations are added to cells in culture. Three to five days later, cell cultures are inspected for... a new vial of early passage cells. Approximately 2 weeks post-thaw, cell culture supernatant is tested...involves transducing cells with serial dilutions of the lentiviral vector, harvesting cells several days later...see maps section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre... (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445...Production Lentiviral preparations are produced in 293T cells using the 2nd generation psPAX2 and pMD2.G packaging...packaging system developed by the Trono lab . Cell culture medium (OptiPro SFM) containing lentivirus is ...

  18. Donations from Addgene to Yield Answers for Rare Disease Researchers

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    Blog Post
    ...transformation of healthy cells into melanoma cells. They’ll combine induced pluripotent stem cell (iPS) technology...gene appears to play a role in many cell processes involving cell membrane dynamics. When they learned...still no treatment available. Aguilar is a basic cell biologist. He got interested in Lowe Syndrome initially...the gene, it piqued their interest. They obtained cells from patients with the condition to reveal some ...explore the effect of various OCRL1 mutations on cell behavior in greater detail. The goal is not just...now plan to use CRISPR-Cas9 technology to create cell lines and/or mouse models carrying different SOX10...

  19. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ...to infect and replicate in host cells, but they are susceptible to cells’ defense mechanisms. The use of... transposon; the plasmid is then degraded by the cell.4.) The transposase creates a double strand break...improve its gene insertion efficiency in primary cells. In 2009, Zsuzsanna Izsvak’s lab screened many SB...tranposase variants for hyperactivity in mammalian cells. Their efforts resulted in a new and improved SB...of antibiotic-resistance genes conducted in HeLa cells). In their Nature Genetics paper, the lab showed...allows the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest...Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol J. 2015 Feb 4. doi: 10.1002/biot...

  20. Plasmids 101: Antibiotic Resistance Genes

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    ... out of 10,000 cells on average! Without some means of quickly determining which cells successfully received...detail selection in non-bacterial cells such as yeast or mammalian cells.   Name Class Mode of Action...Inhibits cell wall synthesis Bactericidal 100-200 ug/mL Carbenicillin beta-lactam Inhibits cell wall ...the bacterium's perspective – a plasmid-containing cell must replicate the plasmid in addition to its own...easily detect plasmid-containing bacteria when the cells are grown on selective media, and provides those...kill bacteria (bactericidal) or slow growth/prevent cell division (bacteriostatic); however, the distinction...
Showing: 181 - 200 of 819 results