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We narrowed to 657 results for: cell;

Showing: 241 - 260 of 657 results

  1. Plasmids 101: Origin of Replication

    Type
    Blog Post
    Published
    Nov. 11, 2020, 2:23 p.m.
    ...to reproduce itself as it must to survive within cells. The replicons of plasmids are generally different...’s sake, we've ignored those used in eukaryotic cells and viruses and focused only on those found in bacteria...example, the pMB1 ori maintains about 20 copies per cell, while pUC – which differs by only two mutations...mutations – will produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard ...positives? Will you have only one plasmid type in your cells at a time? Do you want to make a lot of your plasmid... stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies streaked...bacteria have just reached stationary phase, but the cells have not started to die off. Have any questions...

  2. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...Genome-wide Inhibition Library. DNA and cell amounts After deciding on a cell type, you will need to determine...DNA and how many cells to use for your transformation(s). The amount of competent cells you need total ... generally electroporated into electrocompetent cells to ensure the highest possible transformation efficiency...efficiency. Using Stbl4 or other ultra-high efficiency cells ensures that plasmid diversity in the library is...transformation may vary depending on the particular type of cells you are using, as well as other factors, such as...transformations, such as splitting 400 ng of DNA in 100 μL of cells over four individual electroporations. The specifics...do see recombination, we recommend ensuring the cells used are meant for repetitive sequences (such as...

  3. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...neuroscience where specific tissues, cell types, or even subcellular regions of a neuron need to be targeted...disease, neuroscientists need to be able to probe cells and circuits without disrupting normal brain function...it provides a noninvasive method for manipulating cell activity, especially neuronal firing. What is chemogenetics...drugs. Once activated, chemogenetic receptors hijack cell signaling pathways and can lead to such responses...requires successfully introducing receptors to target cells. Targeted and specific delivery becomes especially...ability of agonists to successfully reach target cells while resulting in minimal off target effects. For...can elicit different responses depending on what cell type it is expressed in. The most common target ...

  4. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...one side of the cell and released again at another side of the cell after intracellular transport without...via individual neurons. Neurons are composed of a cell body, with dendrites receiving incoming information...axon sending information onwards to other neuronal cells. Synapses at the terminals of axons form connections...connections to dendrites of proximal neuronal cells. In the specific example of episodic memory, a subset of... Anterograde tracing outlines neurons from their cell bodies to the terminals of their axons; while retrograde...direction, from the terminals of their axons to their cell bodies. Anterograde and retrograde tracing take...neuronal connectivity a large majority of neuronal cells are labeled overtime due to the continued replication...

  6. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...with enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene offers vectors...reverse transcriptase enzymes, inhibition of host cell proteins, and modifications to the pegRNA. The Britt...shows enhanced editing efficiencies in a number of cell types at different genomic loci — in some cases,... editing efficiencies at different loci in U2OS cells. Image adapted from Yan et al. 2024 under a CC-BY...western blots (Figure 3).      Figure 3: HeLa cells transduced with SARS-CoV-2 nucleocapsid protein ...desirable than having many closely-packed labeled cells. Plus, Voltron2’s fluorescence is tunable by changing...Neuhaus Genetic modification of microglial immune cells is critical for central nervous system research;...

  7. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...differentiate induced pluripotent stem cells (Chavez et al., 2015) into neuronal cells by activating genes that cause...drug targets by finding genes that cause cell death in cancer cell lines upon activation (Gilbert et al.,...Repression and Activation. Cell 159:647–661 . https://doi.org/10.1016/j.cell.2014.09.029 Konermann S, Brigham...and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/10.1016/j.cell.2014.09.039 Wong N, Liu W,... that over-express coding sequences from a given cell type or organism, have been used in a similar manner...many times, the isoform(s) common to a particular cell type are unknown or not readily available. By activating... et al., 2015)) are good choices across multiple cell lines (HEK293T, MCF7, U2-OS, Hela, N2A, 3T3) and...

  8. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    Published
    April 10, 2018, 12:56 p.m.
    .... RANbodies are expressed in 293T cells and are secreted from cells due to their IgGκ signal peptide sequence...-to tool for detecting a protein of interest in cells and tissues. Although antibody production is well... sequence, making them easy to harvest from cell culture media. Their His tags allow for further purification...RANBody While all three reporters stain cultured cells and tissue sections, some reporters are better suited... signal amplification of HRP Staining cultured cells and tissue sections Amplifying weak signals of fluorescent...Labeled anti-HA or -Myc antibody Staining cultured cells and tissue sections When detected with a labeled...Labeled Anti-Chicken IgY antibody Staining cultured cells and tissue sections Great for multicolor staining...

  9. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ...Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System.” Cell 155 (2013): 1379-1491. Pubmed...of 6 chromosome-specific loci in the U20S cancer cell line./ Each color represents a gRNA targeting a ...localization and movement of this locus in live cells! This technique was developed in the lab of Addgene...et al. to track telomere dynamics throughout the cell cycle. To optimize the signal to noise ratio of ...sophisticated tracking of multiple chromosomal loci in live cells. However, they note that certain improvements could...repression and genome editing. Ready to make your cells a little more colorful? CRISPRainbow is available...Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. USA 112(10) (2015): 3002...

  10. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    Published
    Aug. 5, 2021, 12:15 p.m.
    ...sensitive GFP2 (roGFP2)-based biosensors for live cells. His work focuses on the enzyme hydropersulfide-... understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: Micrographs...Micrographs of the biosensor expressed in the human cell line, Hep3B. From left to right: Mitotracker Red, GFP...studies opioid use disorder. Her goal is to identify cell populations in the brain that contribute to the ...will explore the connectivity patterns of these cells and the functional contribution to a variety of ...administration and withdrawal to explore how these cells may encode the negative affect associated with drug...library of mutant sensors expressed in a mammalian cell line engineered to be electrically excitable. The...

  11. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ... defined population of cells (e.g., a neuron subtype) and specific subcellular compartments for measurements...calcium concentrations and transients in living cells and organisms. Unlike chemical indicators, these...neuron-specific expression (hSyn promoter) and pan-mammalian cell expression (CAG promoter). An improved version named...neuron-specific, astrocyte-specific (GCaMP6 only) or mammalian cell expression, and compatible with constitutive or ...measure changes in calcium specific to the neuronal cell body or the axon (GCaMP6 only). The genetic elements...long-term expression and integration within the target cells, ensuring more consistent and reproducible results...calcium sensor to specific tissues or even distinct cell subtypes for efficient expression and downstream...

  12. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    Published
    Nov. 10, 2015, 3:30 p.m.
    ...that inntags IT5, IT6, and IT10 expressed well in cells and did not form undesirable aggregates. Using budding...tags. Impressively, IT6-GFP did not reduce yeast cell size or viability compared to untagged GFP overexpression...multiple other proteins. Again, little to no change in cell morphology was observed with IT5 and IT6 fusions...and HA tags had especially detrimental effects on cell size and growth rate, indicating that they compromised...correlation for protein localization in mammalian cells." Nature methods 10.4 (2013): 315-323. Pubmed PMID...

  13. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...reach essentially every cell or else small pools of antibiotic resistant cells could repopulate the environment...strategies are nonspecific - they harm any bacterial cell without a resistance gene, allowing resistant bacteria...antibiotics target one of three essential processes: cell wall synthesis, DNA replication, or protein synthesis...eliminated from the bacterium, which may also induce cell death. CRISPR also enables increased specificity...Treating a mixed population, represented here by cells of different colors, specifically targets bacteria...bacteria. Targeting a genomic sequence results in cell death. Plasmid targeting may result in plasmid loss...and antibiotic resensitization, or may also cause cell death. Image from Beisel et al., licensed under ...

  14. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...choose cell lines that are highly transfectable.  The amounts of each plasmid transfected into cells are ... polymerase gene or using cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes...capable of being replicated and transcribed in the cell when the appropriate viral proteins are provided... RNA-dependent RNA polymerase into the infected cells. The EBOV polymerase replicates the viral genome...on separate plasmids and are co-transfected into cells alongside a plasmid encoding the minigenome. These...the minigenome) are efficiently transfected into cells. All of these components are necessary to efficiently... RNA polymerase also needs to be present in the cells to transcribe the viral mRNAs and minigenome. This...

  15. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...orthologs that display cleavage activity in mammalian cells. Like Cas9 nucleases, Cpf1 family members contain...the PAM. With this information, they turned to a cell culture system to see if any Cpf1 nucleases would...would exhibit in vivo activity in mammalian cells. From 16 diverse Cpf1 candidates, Zetsche et al. found...particularly helpful for targeting non-dividing cells, which are difficult to modify through homology-... Cpf1 orthologs known to be active in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing... (RNPs) failed to induce off-target editing in a cell culture model. The application of Cpf1 to genome...RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227. Find plasmids from...

  16. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    Published
    June 4, 2019, 1:02 p.m.
    ...non-growing cells are orange, slow growing cells are orange-green, and fast growing cells are green. A...fluorescence. Cells expressing the library are then sorted by fluorescence-activated cell sorting into ...population. Last, Bernach will simultaneously monitor cell cycle and growth rate using TIMER and an unstable...reporter under the control of promoters that control cell cycle gene expression.  Additional Resources on...

  17. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    Published
    April 5, 2022, 1:20 p.m.
    ...cryopreserved hybridomas, immortalized cells created by fusing an activated B cell that produces the antibody of...of interest with a myeloma cell. This RNA was then used to generate cDNA, which  was used as a template...of hybridoma generated mAbs is that if the frozen cell line becomes non-recoverable, the stock of the mAb...hybridoma stocks, without having to grow up the cells. This RNA could then be used to generate functional...immunofluorescence-immunocytochemistry (IF-ICC.) This method was chosen because cells transfected with a plasmid to express the target... could be screened in a high-throughput manner. Cells were labeled with both a hybridoma generated mAb...

  18. Hot Plasmids - January 2023

    Type
    Blog Post
    Published
    Jan. 31, 2023, 2:15 p.m.
    ...mCherry for FACS sorting of the weaponized T-cells (CAR-T cells) - and then delivering them to a tumor - ...effectively as standard CAR-T cells, but it reduces the off-target effects caused by T cell migration.  CAR expression...reducing the on-target off-tumor activity of CAR–T cells was recently devised by the Yingxiao Wang lab.  ...above can be made for expressing your favorite T-cell receptor, or perhaps any other tool that can benefit...  Wu Y, et. al. Control of the activity of CAR-T cells within tumours via focused ultrasound. Nat Biomed...

  19. 5 Great Apps for Lab Life

    Type
    Blog Post
    Published
    Sept. 22, 2016, 2:30 p.m.
    ...’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new...spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find an app that could...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...tools are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution...educational section. This section holds the Gibco Cell culture manual and basic instructional videos. The... Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England BioLabs...alternative, similar apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme...

  20. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...human T cells and hematopoietic stem and progenitor cells (HSPC)s to ~40% expression in the K562 cell line...gene to airway epithelial cells in mice. This vector transduced ~10% of cells, similar to the transduction... then independently packaged into an AAV. When a cell is co-infected with both part A and part B AAVs,... always.  The combination of serotype and tissue/cell type could have a big impact on the rate of transgene...since both of these variables influence whether cells will be co-transduced with multiple copies of both... followed by the 3’-end of the transgene. When a cell is transduced by both viruses, if the AAV genomes...optimal sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview...
Showing: 241 - 260 of 657 results