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We narrowed to 813 results for: cell

Showing: 301 - 320 of 813 results

  1. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...activity. These cells, presumably the cell you want, cannot break down X-gal and are white. Cells that don’t...killing off a portion of the cells like in a selection. Instead, all cells survive and you will need to... the plasmid you’re looking for out of the many cells that don’t. One way to begin the search is by using...using selection strategies, where only cells that have gained or lost a specific gene survive (ex: antibiotic... plates containing X-gal allows you to identify cells that took up the vector and distinguish between ...

  2. Hot Plasmids - February 2022

    Type
    Blog Post
    ... including live-cell imaging, single-molecule tracking, immunolabeling of fixed cells, Western blotting...possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular Cell. 2021. https://doi.org...well-folded scFv scaffold to improve its stability in cells, and use it in a variety of applications. This set...frankenbodies to amplify the signal.  Liu et al. Genes Cells. 2021. https://doi.org/10.1111/gtc.12893, Zhao et...Cas12a, creates challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature...endogenous genes. It is highly specific in mammalian cells without significant off-target effects, yet small...Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized genetic screening technique ...

  3. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ...with) the cells’ own natural translation machinery; these are often incorporated into the cells via plasmid...published, followed soon by GCE done in mammalian cells (Shandell et al., 2021).   How it works If you’...is usually added to the media for uptake by the cells. Second, you’ll need tRNAs and tRNA synthetases ...biosynthetic pathway for the ncAA integrated into the cell’s genome, instead of incorporation via plasmid or...Theoretically, if you put all these together, the cell will produce a protein with a site specifically ...techniques are straightforward, the systems – the cells growing and producing the proteins – are not, and...component of the experiment system (including media, cell type, and growth temperature) may need to be individually...

  4. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ...through programming cells. In the context of therapeutics, if you want to program a cell to identify and ...translational activity a cell has and control how much it taxes the resources of a cell. The objective is to... to the regulatory networks within E. coli host cells. Image from Nielson and Voigt (2014). ...and target a malignant cell, then you’d need the ability to turn on genes at different times. It requires...being used. One thing is that Cas9 is toxic. When cells carry CRISPR circuitry, it can slow their growth...chemicals as sophisticated as those that living cells and animals make. That will be an outcome of genetic...within the wood. There are different types of cellulose fibers, pattern, shape. All of that is controlled...

  5. Working with Nuclear Receptors

    Type
    Blog Post
    ... that are diffused or actively transported from cells, traverse the body through the bloodstream, and ...corepressors depending upon a context defined by ligand, cell type, and promoter, organizing the expression of... coactivator proteins individually contribute to cell regulation in a coherent manner. Because coregulators...that coactivators may be integrators of multiple cell signaling systems. Activation of membrane receptors...sense the impact of the total environment on the cell. The NURSA Consortium Now in its third phase of ...purification proteomic profiling of coregulators in HeLa cells was the first comprehensive analysis of the interactome...executioners of transcriptional regulation. Molecular Cell 27, 691-700. (PubMed) McKenna, N. J.and O'Malley...

  6. Brain Armamentarium

    Type
    Collection
    ...Collection The Armamentarium for Precision Brain Cell Access is developing a collection of molecular genetic... reagents to gain access to many different brain cell types. The expanded collection of Armamentarium ...mCherry-tagged channelrhodopsin-2 driven by PV+ basket cell-targeting enhancer Gordon Fishell Viviana Gradinaru...construct expressing dTomato driven by PV+ basket cell-targeting enhancer Gordon Fishell Viviana Gradinaru...mCherry-tagged channelrhodopsin-2 driven by chandelier cell-targeting enhancer Gordon Fishell Viviana Gradinaru...construct expressing dTomato driven by chandelier cell-targeting enhancer Gordon Fishell Viviana Gradinaru...

  7. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...persist over time, stem cell editing would be highly desirable. Should stem cell editing occur, the CRISPR... high percentage of muscle cells throughout the body, especially stem cells 2. Remove any immunogenicity...patient myoblasts; when injected into DMD mice, these cells expressed functional dystrophin. Editing genes in...in vivo is of course much more difficult than in cell culture, but Ran et al. have shown that CRISPR and...be delivered to both cardiac and skeletal muscle cells, where precision editing of the dystrophin gene ... the percentage of dystrophin-expressing muscle cells increases over time, and all three groups confirm...editing. Tabebordbar et al. show that muscle stem cells are modified by this approach, increasing the possibility...

  8. Plasmids 101: Origin of Replication

    Type
    Blog Post
    ...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...

  9. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...systems, and often do not fold properly once inside cells.  Developing the HA frankenbody To combat the issues...previously proven to fold and function in living cells. To make these scaffolds specific to different targets...the tagged protein upon probe binding in living cells.  In their proof of concept experiment the Stasevich...mCherry, 4xHA tagged histone protein (H2B) in U2OS cells. They then showed that the HA frankenbody probe ...that the HA frankenbody is effective in sensitive cells such as living neurons, and can be fused to other...remains bound to HA epitopes for minutes in living cells, meaning that scientists can track the movement ...effectively used to track single mRNA translation in U2OS cells. HA is small, only 9 amino acids, thus it is translated...

  10. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...plant protein TIR1 in non-plant cells, which interacts with the cells’ endogenous ubiquitin ligase, the...translate AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al., 2016...based method to tag endogenous proteins in HCT116 cells with mAID. Using this method, the lab was able to... protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that...the target sites adjacent to an NG PAM in human cells. The increased versatility of the engineered SpCas9... just a 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised of thousands...

  11. Pooled Library Amplifications

    Type
    Blog Post
    ...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...

  12. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...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 ...

  13. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...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...

  15. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...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;...

  16. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...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...

  17. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    .... 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...

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

    Type
    Blog Post
    ...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...

  19. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...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...

  20. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ... 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...
Showing: 301 - 320 of 813 results