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We narrowed to 872 results for: cel

Showing: 221 - 240 of 872 results
  1. The Importance of High Titer for AAV Transductions

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    ...Making contact with a cell Although an individual AAV particle’s ability to infect a cell is dependent on its...construction and the cell it’s infecting, getting these particles to infect cells is also very titer-dependent... particle to infect a cell, it has to come into physical contact with that cell. Since there is a limited...of surface area on each cell, the way to get more AAV in contact with the cell is to increase the AAV ...between the cell and any individual AAV particle is rare. But simply getting into the cell may not always...particles that have entered the cell now have to travel up the axon, through the cell, and potentially spread ...particles per cell, you may not be able to detect AAV-mediated gene expression in a target cell. Practical...
  2. CRISPR 101: Mammalian Expression Systems and Delivery Methods

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    ...Calcium phosphate Cells Transformed cell lines Transformed cell lines Transformed cell lines Cargo Nucleic...identify and enrich positive cells or a selection marker to generate stable cell lines. Transient or stable...and enrich positive cells. Likely contains a selection marker to generate stable cell lines. Packaging and...wide variety of mammalian cell lines. Useful for difficult to transfect cell types and can be used in ...protein are degraded within the cell.   Delivery methods for mammalian cell lines As mentioned above, the...components into your target cells. Delivery of these components into mammalian cell lines is quite broad and...Electroporation / Nucleofection Microinjection Cells Many types Single cells Cargo Nucleic acids or protein Nucleic...
  3. Viral Vectors 101: Viruses as Biological Tools

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    ...Non-Dividing Cells Transient Gene Expression Low Tropism for Mouse Cells and Hematopoietic Cells Low Packaging...fuses with the target cell’s membrane, allowing the viral particle access into the cell. The viral envelope... of cell the virus will target and fuse with. In nature, this determines which organisms and cells they...viruses can be engineered to be cell type specific. Once inside the host cell, the virus unleashes its cargo...wide range of cell types that are typically ‘difficult to transfect’, such as primary cells. They can also... them to easily deliver their “cargo” to target cells and organisms, and when that cargo is designed to...Viruses as biological tools In nature, viruses infect cells and organisms (which we refer to as hosts) via viral...
  4. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

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    ...genetic access to cortical cell types. Cell. https://doi.org/10.1016/j.cell.2025.05.002 Hunker, A. C.,...Although all cell types share the same DNA sequence, not all enhancers are functional in every cell. Differential...expression in a cell type-specific manner also makes them useful tools for targeting those specific cell types ...Brain Map portal, and cell science-focused tools at the Allen Institute for Cell Science. There are also... primate neocortex. Cell Reports, 34(13), 108754. https://doi.org/10.1016/j.celrep.2021.108754 Panigrahi...generate highly specific tools to target different cell populations and regions of the brain. This has culminated... end_module_block %}   Using enhancers to target cell populations Enhancers are non-coding sequences of...
  5. 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...Fluorescent proteins in cellular organelles: serious pitfalls and some solutions." DNA and cell biology 32.11 (...-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...
  6. Hot Plasmids and Viral Preps - March 2021

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    ...Winslow’s lab to label cells after cell-cell interactions. Using G-baToN, “sender” cells express GFP on the... and “receiver” cells express an anti-GFP nanobody on the cell surface. When the cells interact, GFP is...the receiver cell, and the receiver cell turns green. This system can be used in many cell types including...engineering with single-pot assembly Tracking cell-cell interactions New auxin inducible degron system....brgcm3sw     New system developed to track cell-cell interactions By Jennifer Tsang GFP-based Touching...including endothelial cell-pericyte, neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic...of the interaction between the sender cell and the receiver cell. Image from Tang et al., 2020.   ...
  7. Rabies and Neuronal Tracing

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    ...it first infects one cell (the “starter cell”). It then replicates within that cell, and nascent RABV cross...connections to the starter cell, RABV was modified to only infect user-defined starter cell types, as well as ...dictate infection of target cells through their interactions with target cell surface receptors (Callaway...Callaway 2008). Based on this, cell type specificity of the starter cell can be achieved by using EnvA-pseudotyped...studied biological processes, like how our stem cells are housed in specific locations and give rise to...Tracers Due to their ability to efficiently enter cells and deliver a variety of cargoes, genetically engineered...replicate, and for a virus to cross a synapse from one cell to another, it must replicate. Modified Rabies Virus...
  8. Improving Transduction Efficiency with Exosome AAVs

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    ...the producer cells and purification protocols often describe a process that begins with cell lysis to release...particles is able to infect more cells (as demonstrated in different cellular environments such as retina ... steps to separate cell culture supernatant containing exo-AVVs from producer cells, and then to enrich...Exosomal vesicles secreted from producer cells also contain cellular macromolecules (e.g. proteins, nucleic...effectively target less accessible cells, such as brain tumor stromal cells in a mouse model of glioblastoma...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production... to be spontaneously released from the producer cells into the supernatant during the production phase...
  9. Performing In Vivo CRISPR Screens Using the FITS Approach

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    ...activated T cells, precluding analyses of T cell differentiation beginning with naïve T cells. The Framework...screens have been used to interrogate T cells, an important immune cell type, in more complex and physiological...naïve T cells. More broadly, this system can be used to perform in vivo screens in other cell types. A...screens to assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with...FITS In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However...screens cannot faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that...Framework for In vivo T cell Screens (FITS) approach (Milling and Markson et al., 2024), developed and deposited...
  10. Using AAV for Neuronal Tracing

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    ...one side of the cell and released again at another side of the cell after intracellular transport without...diffusing in the extracellular space; they also have low immunogenicity and cellular toxicity. AAV vectors...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...brain tissue is injected with compounds that intracellularly spread away from the site of application and... Anterograde tracing outlines neurons from their cell bodies to the terminals of their axons; while retrograde...
  11. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    ...unstable, meaning that the cells must continually produce antitoxin to avoid cell death. In the case of plasmids...addiction systems” require dividing cells to retain plasmids - if a cell does not inherit plasmid due to ...will be degraded leaving the cell to be killed by the more stable toxin. Cells only survive if they retain...10% of cells while plasmids containing the plasmid addiction system were retained in 75% of cells. They...consist of a toxin which can affect a variety of cellular processes and an antitoxin that suppresses the...designated this fragment as the ccdB region (for “coupled cell division”). The encoded toxin-antitoxin system consists...cleaved complex. This results in DNA breakage and cell death. Conversely, CcdA binds to and blocks the ...
  12. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    ...particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor and sometimes ...transduction efficiency, and is an excellent choice for targeting cardiac cells (Bish et al., 2008). It can ...-associated viruses. Cell, S0092-8674(25)00692-0. https://doi.org/10.1016/j.cell.2025.06.026 Halbert, ... Therapy: An Overview. Cells, 12(5), 785. https://doi.org/10.3390/cells12050785 Kaludov, N., Brown, K....tissue tropism, allowing them to target particular cell types. Figure 1: Diagram of AAV components...key that lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic...multiple coreceptors, which may be present on different cell types and sometimes vary between target species ...
  13. CRISPR/Cas9 FAQs Answered!

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    ...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....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. ...pluripotent stem cells differing exclusively at two early onset Parkinson Point mutations. Cell, 146(2), 318...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...
  14. Plasmids 101: Control Plasmids

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    ...untreated cells—but can we be certain? Do we know whether the plasmid delivered into the cells successfully...amount of DNA transfected into the cells and the ability of the cells to express the protein.   In analysis...Gene X in backbone Y) was transfected into human cells. From this result one could simply conclude that...didn’t work, as the expression level of Gene X in cells treated with Plasmid A is similar to the expression...or transfection process) may have on the target cells. One transfection control is an empty vector control... itself has any cytotoxic effects on the target cells. Another type of transfection control is an internal...plasmid or transfected into a separate well of your cells. Regardless, the amount of reporter protein activity...
  15. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    ... is technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to... Dyes that can pass through cell membranes can prove useful for live-cell imaging. Experimental context...Additionally, cell permeability is a critical factor, as it influences the dye's ability to enter cells without...enabled the visualization of both organic tissues and cell cultures, opening biological interiors to many inquisitive...fluorophores to enable highly specific visualization of cellular components, typically a protein of interest. Fluorescent... dyes/ligands Effects on protein function and cellular components: It's important to assess how dye-conjugation...affects protein function and its propensity to bind cellular components. Some dyes target specific organelles...
  16. Genome-wide Screening Using CRISPR

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    ...heterogeneous population of mutant cells, with each cell or set of cells containing a mutation in a different...separately.  Mutant cells are enriched using either drug selection or fluorescence-based cell sorting and screened...mutant cells can be used in drug screens to identify genes that confer drug-resistance. Mutant cells are ...generate lentivirus (B). Cas9-expressing cells or wild-type cells are treated with lentivirus containing...plus Cas9 to generate the mutant cells, respectively (C). Mutant cells are screened (D) and "hits" are ...lead to skewed results. Selecting a cell type - Theoretically, any cell type can be used in a CRISPR screen...requires a massive amount of cells as starting material. Therefore, cell types that are of low abundance...
  17. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    ...this approach is limited to protoplast cells (plant cells whose cell wall has been removed). Another common...expression is dependent upon the cell type in question. In plant cells, gRNA expression is achieved by ... shown to be excellent alternative genome editing technologies especially in human cells. In contrast ...are necessary to enable genome editing in plant cells.   This blog post will present a general overview...are necessary to use the CRISPR system in plant cells. Like other model systems, expression of S. pyogenes...gRNA) is sufficient to modify the genome of plant cells. The structure of the gRNA (composed of a ~20 nucleotide...increase translation in a particular plant species or cell type (Belhaj et al., 2013). Nuclease dead Cas9 (...
  18. Viral Vectors 101: Optogenetic Tools

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    ...pulses to the subject/cells expressing the optogenetic tool causing a change in cellular activity or gene ...receptors (GPCR) regulate cellular function through multiple intra-cellular signaling cascades. Optical... in the extracellular domain which activates the native signaling cascade of the intracellular part of...Neuroscience. Frontiers in Cellular Neuroscience, 15, 778900. https://doi.org/10.3389/fncel.2021.778900 Mattis...positively charged ions to rapidly pass through the cell membrane and initiate an action potential - a very...have developed many optogenetic tools to control cellular activity with exquisite precision using only light...both the membrane potential and a host of other cellular functions. Photoswitches are engineered molecules...
  19. Viral Vectors 101: AAV Variables That Matter

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    ... where many different cell types, or even different phenotypes of the same cell type, are intermingled...postsynaptic cells when used at very high titers (Zingg et al., 2017), which can lead to cells in far away...Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org/10.3390/cells12050785 Murlidharan, G., Samulski...Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org/10.3390/cells12050785 Kanaan, N. M., Sellnow...experimental conditions and measurements; (2) the cells you’re targeting; (3) how much expression is needed... capsids, which determine how a virus enters the cell. Different serotypes utilize a number of different...co-receptors, and other mechanisms to gain entry into the cell (Dudek et al., 2018; Korneyenkov & Zamyatnin, 2021...
  20. 15 Hot Plasmids from 2017

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    ... of cell types in the nervous system. While certain cell types, such as principal excitatory cells and...neuronal cell types involved in neurodegenerative diseases, the Cole Lab selected promoters for cell types...yield of 0.70), mScarlet-I with T74I (accelerated maturation in cells compared to mScarlet, fluorescent lifetime...with edited “pools” of cells, eliminating the time-consuming step of single-cell isolation. Agudelo et ...gene. Koo M, et al. Cell Systems. 2016. PubMed PMID: 28189581 Peters J, et al. Cell. 2016. PubMed PMID:...generation Deriving iPSCs (induced pluripotent stem cells) through the expression of transcription factors...the established model for reprogramming somatic cells. A common method for generating iPSCs to express...
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