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We narrowed to 657 results for: cell

Showing: 161 - 180 of 657 results

  1. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    Published
    Jan. 30, 2014, 3:29 p.m.
    ... 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...

  2. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...specific cell types (populations) within neuronal networks in living animals. To accomplish such cell specific...individual cells. With this technique, a Neuroscientist can see the full dendritic arbor of a single cell with...Cre-mediated possibilities for site specific (and often cell type specific) control of DNA recombination and ...30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular evolution...revolutionize neuroscience Manipulation of specific cell types in neuronal networks Double inverted vectors...combination with mouse lines that express Cre only in the cell type of interest (Atasoy et al. 2008; Cardin et ...single neuron and its synapses in the living brain? Cells of a specific brain area can be transfected by in...

  3. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ... brain, robust single-cell labeling is necessary to distinguish individual cells. The Brainbow-1.0 construct... May 5. PubMed. Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish... 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing in other systems, such...Brainbow highlights the trend in biology towards single-cell resolution imaging. Sanes and Lichtman endeavored...door for immunostaining analysis. To achieve even cell labeling, they generated farnesylated derivatives...fluorescent proteins, which are directly trafficked to cell membranes. Membrane trafficking of farnesylated ...Brainbow mice to determine the number and type of cells in which the construct is expressed. The photostability...

  4. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    Published
    Nov. 19, 2019, 2:08 p.m.
    ...OptoNBs to control cell signaling The second application for OptoNBs takes us into cells: light-based control... regulation of endogenous signaling activity in cells.  Let’s shed some light on the key components of...their use for protein purification and controlling cell signaling. Key components of OptoNBs OptoNBs have...can bind, it co-localizes with its target at the cell membrane and when the OptoNB can’t bind, it localizes... over-activated in cancer due to its involved in cell proliferation and differentiation.  Since Ras is...They expressed this OptoNB-SOS fusion protein in cells that expressed membrane-localized mCherry. With ...the nucleus where gene expression changes lead to cell proliferation and differentiation. The lab confirmed...

  5. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (5 December...CRISPR in adult stem cells has huge potential, because the genomes of adult stem cells in organoid cultures...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (5 December 2013). 8. Gerald Schwank...homologous recombination in cultured intestinal stem cells of patients with cystic fibrosis (8).   The difference...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis....is a big advantage over induced pluripotent stem cells, which need to go through a phase of crisis where...B. 2021. “CRISPR/Cas gene therapy.” Journal of cellular physiology.  https://doi.org/10.1002/jcp.30064...

  6. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...Virus (MoMLV, MMLV, MuLV, or MLV) or Murine Stem Cell Virus (MSCV) genomes whereas lentiviral vectors ...things) gene delivery, generating pluripotent stem cells, and developing gene therapies for monogenic diseases...vectors, their infectivity is limited to dividing cells. Find tips on using gamma-retroviral vectors in ...are able to transduce dividing and non-dividing cells. More detailed descriptions of the lentiviral elements... to express siRNAs of their choice in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi...-optimized Cas9 under the control of Murine Stem Cell retrovirus promoter for mammalian gene knockdown...96 retroviral plasmids (based on the Murine Stem Cell Virus, MSCV) each with a unique 6 nucleotide barcode...

  7. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...vectors (right) are transduced to target cells. Within target cells, Cas9 machinery is transiently expressed...in order to facilitate its entry into the target cells (Beatty et al., 2012). These features make AdV an... complexes into transformed and non-transformed cells yielded rates of targeted mutagenesis similar to...nuclease-induced gene disruption frequencies in the various cell types ranged from 18% to 65% (Maggio et al., 2014... leads to a vast majority of AdV-modified human cells being subjected to scarless homology-directed genome...therapeutically relevant, non-transformed mammalian cells. These viral vector systems also work equally well... dividing and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral ...

  8. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ...for studying the bacterial cell cycle, asymmetric cell division, and cellular differentiation, but it takes...crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-based DNA demethylation Optogenetic repressors...Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image...lab repressed ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and...piggyBac transposon system for human pluripotent stem cells Lindy Barrett’s lab developed new fluorescently ... of multiplexed gRNAs in human pluripotent stem cells. Previously, CRISPRi or CRISPRa depended on continued...KRAB and dCas9-VPR for quantifying and tracking cells that express both the dCas9 construct and the fluorescently-labeled...

  9. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    Published
    June 24, 2014, 3:59 p.m.
    ...luciferase assays require cell lysis as the most efficient means to disrupt the cell membrane and deliver ...miRNAs for effectiveness. Analyze Cell Signaling Pathways Known cellular signaling pathways can be investigated...luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays have since become a gold standard... serves as an indicator of transcription within cells, where detection of the reporter protein or its ... into the desired mammalian, plant or bacterial cells. Expression of luciferase as the reporter gene is...can permit measurements of luminescence from live cells. Monitor miRNA knockdown Expression of luciferase...codon, which is then transfected into the desired cells. A hybrid mRNA transcript containing luciferase ...

  10. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ... engineer cells for specific purposes. Unfortunately, plasmid copy numbers vary within cell populations...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes.... When cells are dividing rapidly, multiple rounds of replication are initiated before the cell has divided...growth rate can all impact plasmid copy number in a cell. One can attempt to eliminate fluctuations due ... pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered against the effects... to stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold...fold (for p15a, R6K, and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro...

  11. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...illumination of neighboring cells evokes action potentials (APs) in patched cells expressing wild type CoChR...techniques for live-cell imaging of the dynamic changes in biochemical activity within the cell. Their Fluorescence...28646204. soCoChR: A novel opsin that enables single-cell optogenetics Article contributed by Meghan Rego ...wild type CoChR, soCoChR localizes primarily to the cell body with very little signal detected along the ...CoChR, these APs are absent in soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish...precise mapping of neuronal networks at the single-cell level. Various soCoChR plasmids described in this...strain to assay for functional decaffeination in cells. The ΔguaB E. coli are missing an essential enzyme...

  12. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ... expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown... complex. Not performed in a native cell context. For AP-MS, cells are lysed before the bait protein is...and interacting proteins in mammalian cells. The Journal of cell biology. PubMed PMID: 22412018. PubMed...against PPIs that aren't stable outside the native cell context. The Split-BioID method Enter Split-BioID...tagging of proteins in both BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein...proteins that interact with the bait in a native cell contact. Other PPI methods may require protein interactions...interactions to remain stable (or to be stabilized) during cell lysis and lysate fractionation. Important considerations...

  13. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ... do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were transferred...functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside the...a wide range of cell types including but not limited to muscle, liver, and brain cells. There are a multitude...chain (c) and is characterized by a lack of immune cell development and function. To date, gene therapy ...that can deliver gene therapies directly to the cells that need them. Adeno-associated virus (AAV) – based...delivery of nucleic acid cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery...expression over many years in postmitotic, long-lived cell types. AAVs in the clinic Thus far, AAV vectors...

  14. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...large amounts of carbohydrates that can inhibit the cell lysis process. To ensure that lysis is complete,...strains adapted for protein purification use the cell’s resources to produce high levels of protein, not...difference? In the simplest of terms, more volume = more cells = more plasmid. Plasmids can vary in copy number...of plasmid copies supported within each bacterial cell in your culture. Copy number is partially specified...richer media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield... supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific...glucose (in moderation) may also serve to increase cell density and plasmid yield. Start with a single, ...

  15. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...same: Deliver CRISPR library to cells of interest. Select for cells of interest (positive or negative...infect specific cell or tissue types. Using serotypes that have a known tropism for your cells of interest... a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing cells for long...mice are involved. Ex vivo starts with transducing cells with the CRISPR library in vitro before transplanting...interest — for example, AAV8 for kidney cells — can help improve infection rates in your experiment. ...AAV vectors, enhancing their affinity for specific cell and tissue types. The combination of serotypes and...integration sites seem to only occur in the target cells or tissues. AAV also shows improved transduction...

  16. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... 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 two weeks post-thaw, cell culture supernatant is tested...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...preparations ("-LV” catalog items) 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 first...modified from Wang et al., 2018). Briefly, 293T cells are transduced with serial dilutions of a lentiviral...

  17. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...number of cells have been edited. For many experiments, the end goal is to generate a clonal cell line, so...designed your gRNA and introduced it into your target cells with Cas9. Hooray! Now it’s time to make sure your...blog post we’ll explain how to verify that your cells were appropriately edited for your desired mutation...important to know a few things about your target cells and the desired edits. First, you will need to know.... Validating knockout mutations When a knockout cell line is desired, any mutation that introduces a ...input is sequencing trace files of WT and edited cell lines. Then decomposition analysis is performed ...both an unedited population and your Cas9-targeted cells. Simply upload both trace files to the TIDE tool...

  18. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...scFvs and T cell receptors and are expressed on T cells to make CAR T cells. The extracellular scFv portion...Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific antigen, it brings T cells to a ...scFv part of the receptor can activate T cells to kill cancer cells. scFv’s small size also allows for their...recognizes an antigen of interest while the intracellular T cell receptor portion of the CAR facilitates ... display libraries screen antibody genes from B cells of immunized animals, but it's possible to avoid...also allows for screening scFv generated from the B cell antibody genes from diseased or vaccinated humans...produce scFvs in mammalian, yeast, plant, and insect cells, they are most often expressed from a plasmid in...

  19. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    Published
    Jan. 11, 2017, 8:47 p.m.
    ...both SpCas9 and your gRNA in a variety of mammalian cell types. Purpose  SpCas9 This plasmid expresses Streptococcus...the Zhang lab has tested in a variety of mammalian cell lines. Cas9, of course, is the key component of ... puromycin resistance gene enables selection of cells with the plasmid post transfection. The 2A "self...can be used to transfect a variety of mammalian cells and can be modified for viral trasduction (see the...resistance gene that made it less effective in certain cell lines. This point mutation has been corrected in... your gRNA of interest in a variety of mammalian cell lines. Alternative forms of this vector contain ...Common Ways to Deliver CRISPR Plasmids to Mammalian Cells Resources on the Addgene Website Visit our CRISPR...

  20. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...transducing the cell type under study. Many AAVs will show significant expression in a variety of cell types, ...optogenetics as tools to investigate the roles of certain cell types in locomotion. We have relied heavily upon...deliver chemogenetic/optogenetic constructs into our cells of interest. Although they have a small packaging...of serotype (which determines which tissues and cells an AAV can infect) can influence an AAV’s value ... tissue? Location (2): Where in the tissue/which cell type? Location (3): Traveling from the original ...AAV5, AAV6, AAV9 Pancreas AAV8 Photoreceptor Cells AAV2, AAV5, AAV8 RPE (Retinal Pigment Epithelium...-vectors/aav/aav-guide. Where in the tissue/what cell type? In the CNS, for instance, there are marked...
Showing: 161 - 180 of 657 results