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We narrowed to 762 results for: cell

Showing: 201 - 220 of 762 results
  1. Viral Vectors 101: What is a Virus (and a Viral Vector)?

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    ...hijacking the machinery of the cells they infect. Once they enter a cell, they treat their viral genome...genome as a “payload” that is delivered to the cell. The cell then uses its own machinery to replicate the...replication process whereby one cell copies itself and “splits” into two cells. Viral vectors Viruses are ...virus. From one, come many After a virion infects a cell, the single particle creates many copies of itself...millions of new copies, which are released from the cell like baby spiders from an egg sac. (Okay, some enveloped...lysing, but they do generally still kill the infected cell.) This is in contrast to bacteria and eukaryotes...means the vector can deliver a genetic payload to a cell but cannot create new particles. As a bonus, this...
  2. Mapping the 4D nucleome with CRISPR/Cas9

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    ...labeled Cas9 to label multiple genomic loci in live cells. While other systems can be used to label loci, ...CRISPR/Cas9’s ease of use and ability to label live cells make this system truly advantageous. This new technique...nuclear organization varies across the life of a cell, or how organization may be altered in disease states...dynamics, and how they change in response to the cell cycle or other stimuli.   To create a colorful Cas9...the signal level provided by SunTag is so high, cells may be imaged under lower illumination settings,...Cas9-based method is a great step forward in live cell imaging of genomic loci. Ma et al. anticipate that...that the technique will be applicable to studies of cell cycle progression, epigenetics and cellular reactions...
  3. An Integrin Antibody Toolkit from IPI

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    ...every cell in the human body, integrins are key to a diverse array of processes, including cell adhesion...tensions before they would enable cells to spread, resulting in cells that elongated and morphed asymmetrically...on the BJ-5ta fibroblast cells, integrin αVβ1 was critical in initiating cell spreading and two other ...Integrins are essential cell surface receptors involved in complex signaling pathways and the linkage...these receptors have proved challenging to study. Cells often express multiple integrins that bind the same...ligand-binding heads and long legs that protrude from the cell membrane, physically connecting the extracellular...is key to strengthening ligand binding, mediating cell adhesion and migration, initiating intracellular...
  4. Mouse Modeling, Part 1: Genetically Engineered Mice

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    ...modified zygotes or embryonic stem cells. Modified embryonic stem cells are injected into a blastocyst which...engineered cells. Similar to transgenic mice, knockout mice are engineered during the embryonic stem cell stage...not all cells in the blastocyst were successfully edited by the injection of embryonic stem cells. Also,...controls the cell types in which the conditional recombination can occur based on the cells in which Cre... interest into a zygote, or into embryonic stem cells that are then injected into a blastocyst. There ...of interest into a zygote or into embryonic stem cells. Non-homologous recombination incorporates this ...targeting vector is electroporated into embryonic stem cells and this will insert the gene at a specific site...
  5. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

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    ...proteins in ten ddifferent cell types, including difficult-to-manipulate primary cells, achieving knockdown ...stable cell lines overexpressing TRIM21, finding that TRIM21 overexpression did not affect cell proliferation...electroporated cells re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines, endogenous...proliferation. These cells also did not contain changes in expression of TRIM21-associated genes, indicating...multiple methods for TRIM-Away delivery. For small cell populations, co-microinjection of TRIM21 protein...protein and the target antibody works well. In larger cell populations, you can deliver these components together...together using electroporation. In these bulk cell experiments, Clift et al. observed no stress or damage...
  6. Plasmids 101: Blue-white Screening

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    ...gets interrupted as shown in cell A, and thus will not complement the cell's β-galactosidase mutation. An...therefore the cell will have a functional β-galactosidase enzyme through α-complementation (cell B). As shown... plasmid-containing cells without using antibiotic resistance. These rely on cell lines susceptible to...without which the cells cannot reproduce to form colonies (used to complement auxotrophic cell lines), or genes...called the α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a...screening process. It does not select only those cells that have taken up a plasmid and thus should be ...white due to successful cloning and not because the cell failed to take up the α-complementation plasmid....
  7. Exploring Applications of the Bioluminescent HiBiT Tag

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    ...kinase essential for cell proliferation signaling. CRISPR knock-in HiBiT-tagged EGFR cells allowed researchers... fusions utilizing bulky tags can disrupt normal cell biology and make it hard to capture true protein...researchers with a means to quickly iterate, work across cell models, and even introduce mutations. This vector-based...and other fluorescent proteins revolutionized live-cell protein detection, their bulk (~27 kDa) can perturb...Bioluminescence detection is a cornerstone of modern cell-based assays and enables ultra-sensitive detection...phototoxicity, perturb biological processes, and compromise cell viability (Laissue et al., 2017). HiBiT overcomes...EVs), edited transcripts could endow neighboring cells with pro-oncogenic traits, facilitating metastatic...
  8. Plasmids 101: Choosing an Antibiotic Resistance Gene

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    ...prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells. The...synthesis), while amp only affects the cell wall synthesis step during cell division. This is nifty because ..., penicillin, and is a beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces...at 37 degrees) can be shortened since the plated cells will have time to produce beta-lactamase prior to...penicillin family of antibiotics. It interferes with cell wall synthesis as its mode of action (sound familiar...culture, but some types of bacteria, such as SHuffle cells, are already resistant to this antibiotic due to...the bleomycin family of antibiotics, which induce cell death by intercalating into DNA and generating DNA...
  9. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    ...numerous cell biology assays. However, there has been a pervasive view that most FPs in a cell will fold...fluorescent fusion proteins in cell biology."Current Protocols in Cell Biology (2005): 21-4. Pubmed PMID...Lee. "Fluorescent proteins: a cell biologist's user guide." Trends in cell biology 19.11 (2009): 649-655...-fusions on quantitative imaging experiments and cell physiology in general. Pitfalls of Fluorescent Protein...quantitative imaging experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable...dynamics of fluorescent protein fusions in live cells in real time. Previously, we have described a number... i.e. positioning of the FP sequence relative to cell compartment targeting sequences, which often have...
  10. Plasmids 101: Screens vs. Selections

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    ...culture cells in the drug of interest at a lethal dose for non-manipulated cells and then assay cells which...selection, you might put cells into a drug-containing media in which only cells which can tolerate the ...selection. A selection is a process in which only cells with the trait of interest will survive in the selected... – that is, you are not specifically looking at cells that can survive. Instead, all manipulations are...off. For a screen, you would individually treat cells with many different drugs and then perform an assay...manipulating or mutagenizing a population (typically cells or small organisms) and then assaying for a functional...For example, a drug screen may be performed on a cell line possessing a patient mutation to assess how...
  11. Supporting Rare Disease Research with the 2020 BeHEARD Awards

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    ... trigger the activation of dendritic cells, macrophages, T cells, and the recruitment of neutrophils. ... says. In induced pluripotent stem cells derived endothelial cells from women who are missing an entire...maintaining the nutrient and ionic balance of the cells,” says Dr Sangeetha Iyer, ACD’s Scientific Advisor...that solute channels typically expressed on other cell types can compensate for or bypass the dysfunction...collection to perform a functional screen in mammalian cells to identify any solute channels that can compensate...be able to validate their findings in mammalian cells with both human and murine targets thus informing...chromosome (TS iPSCs), the lab observed abnormal cell migration and contraction. The lab hypothesized ...
  12. 28 Hot Plasmid Technologies from 2015

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    ...in live cells is that one can now follow movement of two or more targeted loci during the cell cycle or...mammalian cells, yeast, and Drosophila. Synchronization of secretory protein traffic in mammalian cells The... different organelles, a cell cycle indicator for mouse embryonic stem cells and a series of inducible...populations within the same cell. Mitochondrial fusion is typically assayed in living cells by using mitochondrially-targeted... alternative splicing in mammalian cells. Younis, et al. Mol Cell Biol. 2010 Apr;30(7):1718-28. New...yeast cell viability, had less of an effect on GFP diffusion, properly localized in mammalian cells without...fluorescent fusion proteins can visualize: Cell Structure Plasmids: Cell Structure Plasmids: ER mCherry-Sec61...
  13. An Introduction to Adenovirus

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    ... AdVs is dependent on modulation of the host cell’s cell cycle; inhibition of the innate immune system... in the producer cells where there is E1 and E3 gene complementation. In target cells that are infected...death of adenoid cells in vitro (Rowe et al., 1953). While overt death of the targeted cells is less likely...infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(4), 2296....production and/or by genes present in the HEK293T cell line used (which have fragments of the AdV genome...achieves persistent transgene expression in certain cell types (Tatsis et al., 2007), potentially because...viral genomes or otherwise clear the virus from the cell population (Kafri et al., 1998). In fact, adenovirus...
  14. Hot Plasmids - August 2020

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    ...for labeling specific structures within cells. HeLa cells were transfected with plasmids encoding...protein evolution Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system...mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image from Mateo...labeling structures and compartments in mammalian cells By Angela Abitua Genetically encoded fluorescent...or specific localization of biomolecules within cells. However, sometimes these fluorescent proteins might...exactly a structure or protein is located within a cell. The Goedhart lab, Gadella lab, and collaborators...for labeling a list of specific structures within cells. For example, to improve nuclear localization of...
  15. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    ...can often be used in cells that are difficult to transfect, such as primary cells. Using RNPs can also...interest. This is commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9...must be considered when using this method:     Cells must be amenable to transfection or viral transduction...protein in complex with a targeting gRNA, to your cells of interest. Cas9 RNPs are capable of cleaving genomic...single or multi-gene knockouts in a wide variety of cell types, gene editing using homology directed repair...difficulties with protein expression that occur in cells where common eukaryotic promoters (such as CMV or...after transfection, and are quickly cleared from the cell via protein degradation pathways. This makes Cas9...
  16. AAVs CREATed for Gene Delivery to the CNS and PNS

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    ...that cells are labeled with their own unique color; and 2) low levels of labeling so that a cell’s projections...tables 1 and 2, with a cell-type specific promoter or enhancer now allows for cell-type-restricted expression...this plasmid pool are injected into an animal with cell-type-specific Cre, the polyA sequence is inverted... library to infect an animal expressing Cre in a cell type of interest, a scientist can isolate DNA from...(s) of the capsid gene successfully infected the cell type of interest. Repeat rounds of infection and...capsid variants with increased ability to infect the cell type of interest, which, in the case of Chan et ...transduced a higher number of neurons overall (GFP+ cells that also stained for a neuronal marker). Check ...
  17. Creating accessible biology activities in schools with BioBits

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    ...on freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free extracts Typically... into a host cell (through a process called transformation), essentially giving the cell all the instructions...to dispose of the live cells. The alternative is to express the protein in a cell-free manner. This is ...electrophoresis to separate DNA or transforming cells with green fluorescent protein, these activities...transcription and translation. Unfortunately, having live cells in a classroom presents equipment needs, such as...operating the rest of the car as well. Similarly, cell free expression systems contain all the parts needed...handle a living, replicating organism. Furthermore, cell free systems can be freeze-dried into pellets (FD-CF...
  18. Genetic Code Expansion

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    ...applications of genetic code expansion in cell biology . Nat Rev Mol Cell Biol, 13 (3), 168–182. https://doi....expanding the genetic code in bacteria and mammalian cells. Plasmid...which means they work independently of the host cell's natural genetic machinery. The orthogonal aaRS ...while E. coli pairs are orthogonal in eukaryotic cells. Pairs from methanogens like Methanosarcina barkeri... are orthogonal both in bacteria and eukaryotic cells. Many of these sets have been mutated and screened... Once these plasmids have been introduced in the cell, the ncAA provided in the media can be incorporated...match the growth medium, ncAA concentration, and the cell lines used previously. Remember that the orthogonal...
  19. dTAG - You're it!

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    ...dTAG molecule toxicity in new cell lines Treat parental cells (or cells expressing a control -- for example...dTAG system is versatile and can be used in both cell-based assays and mouse models. The dTAG system The...of N- and C-terminal fusions and degradation in a cell line of interest prior to starting knock-in experiments...the FKBP12F36V-fusion chimera of interest. In the cell lines we have evaluated to date, we observed little...different contexts (for example, IZKF1 and IZKF3 are cell-type specific transcription factors with important...selective FKBP12F36V fusion chimera degradation. In the cell lines we have evaluated to date, we have not observed...system was employed to evaluate degradation of ENL. Cell lines were engineered to express ENL-FKBP12F36V ...
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