We narrowed to 1,027 results for: ELL

...infecting most cells, but not all. For example, LDLR expression in hematopoietic stem cells, T cells, and B cells... host cell. Some of these envelope-receptor pairings are broad, allowing delivery into many cell types...delivery to only a few cell types.  If you’re looking for more control in the cell types your lentiviral...protein causes cells that it infects to fuse together and form syncytias that cause the cells to die. Cytotoxicity...Cytotoxicity can also be cell type dependent: if a cell has few receptors for a specific envelope protein...lipoprotein receptors (LDLRs) on a cell surface to endocytose into the cell. The reason VSV-G is so commonly... cells is low. Infection of these cells with VSV-G pseudotyped virus requires the upregulation of LDLR...

...Fluorescent proteins in cellular organelles: serious pitfalls and some solutions." DNA and cell biology 32.11 (...localize to chemically distinct subcellular environments, including the organelles that compose the secretory...numerous cell biology assays. However, there has been a pervasive view that most FPs in a cell will fold...fluorescent proteins for subcellular environment. Currently, she is a SPIRE postdoctoral fellow at the University...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...

...one side of the cell and released again at another side of the cell after intracellular transport without...typically used for the trafficking of organelles, such as mitochondria, as well as macromolecules like actin and...diffusing in the extracellular space; they also have low immunogenicity and cellular toxicity. AAV vectors...tracing has the added benefit of only labelling the subset of infected cells that also express Cre. Another ...requires both antero- and retrograde labelling, synaptic restriction as well as controlled monosynaptic and ...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...

... into cells in the lab. The introduction of foreign DNA or RNA into bacteria or eukaryotic cells is a ...application of an electrical charge to cells, to increase cell membrane permeability and thus transformation...introduce foreign DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral...the genome and stable cell lines, where foreign DNA is incorporated into the cell’s genome and is thus passed...through cell division. You can find all kinds of different lentiviral and AAV plasmids as well as ready-to-use... pokes holes in the cell membrane so DNA can be introduced directly into the cell. Microinjection requires...multiple ways foreign DNA can be introduced into cells including transformation, transduction, conjugation...

...model systems: either human cells or mice. In a recent issue of Cell Stem Cell, Musunuru, Chad Cowan and...done in stem cells, we can differentiate those mutant and wild type stem cells into any cell type relevant... the mutant cell lines that one needs to do disease modeling in stem cells. Addgene: Tell me about the...system for generating human stem cell-based disease models." Cell Stem Cell. 12, 238-251 (7 February 2013...homology-directed repair by flooding the cell with a mutant template and fooling the cell into using that template for...FokI endonuclease domain as well as fluorescent proteins used to sort the cells after the plasmids have been...unique aspects that make it especially well-suited for use in stem cells. (See the complete protocol at StemBook...

... the single cell level), are separated by, for example, cell sorting. The separated cells are subsequently...more than 8x107 successfully targeted cells. Depending on the cells used and the readout, this could be ...My group works on immune cells in different contexts. Most of the immune cells we study are migratory, ...CRISPR modified cells are transferred into an animal and the behavior of the transferred cells is followed...with an easily measureable cellular phenotype (e.g. survival of a cancer cell as a cytotoxic drug is added...genes essential for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683. PubMed PMID:...With these screens, the investigator generates a cell population where all genes are knocked out at the...

...10 µM oligo stock for each well on 24-well plate, co-deliver into the cells with the Cas9/sgRNAs vector...find both single-allelic and bi-allelic cells. Single-allelic cells usually make up the majority in culture...donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example/reference is Ran et al...plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good reference is Yang et al. (2013...tract length in Drosophila. Elliot et al. (1998) suggests mammalian cells' tract lengths seem much shorter...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....

...transgenic cell lines? One of the major uses of the piggyBac™ system is to create stable transgenic cell lines...efficiency is likely to be cell line-dependent, and should be tested in the cell line of interest. Importantly...If the cell’s chromatin structure undergoes significant rearrangement (such as when a stem cell initiates...manipulations in animal models or cells lines, including stem cells, without leaving behind any accessory...applications, including making transgenic cell lines and genetically-modified cell lines. Furthermore, the use of...molecular and cellular biology and brings 10 years of research experience with stem cells to Transposagen's... in Human Pluripotent Stem Cells: Choosing the Correct Path.,” J. stem cell Regen. Biol., vol. 1, no. ...

...to the well (either directly or by binding to an antibody that is coating the bottom of the well), and ...coating the wells to capture antigen from a sample, whereas antigen is bound directly to the wells in a direct... the wells are typically already coated with the antibody. Then, you add your sample to the wells, and...sample are bound to the wells of a plate instead of an antibody. Coating the wells with a purified protein...ELISA well As mentioned above, the measurement of the concentration of proteins in an ELISA well is based...the reaction, turns the solution yellow. The absorbance of each well, which is measured using a microplate... are used to detect proteins within a 96- or 384-well, flat-bottomed plate. The basic principle of an ...

...contaminating your transduction with the packaging cells. Packaging cell debris can be removed either by filtration...-500 g). After removing packaging cell debris, virus can be pelleted by ultracentrifugation (75,000 - ...ultracentrifugation, virus may appear as a white pellet. The pellet can then be rinsed and resuspended in cold... you can reduce the volume of cell culture medium on the packaging cells immediately following transfection...efficiency in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and... the packaging cells after 1–3 days of selection. A lower percentage of surviving cells may be the result...adeno-associated virus transduction of cells." American Journal of Physiology-Cell Physiology 291.2 (2006): C386...

...restoring mCherry expression. Cells can then be sorted using Fluorescence-activated cell sorting (FACS) and the...easily determined by comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry...expressed in cells. mCherry is constitutively expressed to identify successfully transfected cells. C-to-T ...eGFP and mCherry positive cells by the number of only mCherry positive cells (Martin et al., 2019). ...TREE) to purify single base edited cells without the need for single cell isolation and downstream sequencing...fluorescence and cell sorting using FACS which could limit their use across different cell types and systems...sgRNA targeting the GFPACG to the cells. As expected, only the cells that contained the cytidine base ...

... carried by extracellular vesicles (EVs), edited transcripts could endow neighboring cells with pro-oncogenic...observe degradation as well as endocytic trafficking in real time through extracellular-specific and full ...kinase essential for cell proliferation signaling. CRISPR knock-in HiBiT-tagged EGFR cells allowed researchers...Degradation of Misfolded CFTR. Cells, 12(23), 2741. https://doi.org/10.3390/cells12232741  Kamio, T., Kono, Y.,... 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...

...be different for each transduced cell, there’s no telling which cells may harbor these unwanted interruptions...purposes, like creating stable cell lines, delivering CRISPR/Cas9 to cells for genome editing, and introducing... chromosome of the cell. This integrated viral genome is then part of the cellular DNA… forever!  This...viral entry into the host cell.  Transfecting these three plasmids into cells enables the production of...lost through cycles of cell division, though the rate of loss greatly depends on cell turnover. Why use ...expression in non-dividing cells IDLVs can also be useful for transducing non-dividing cells, as they avoid the... transduced cells and quantifying the integrated viral DNA along with a reference cellular gene. When ...

...Mammalian Cells pCMV-DnpK dinitrophenyl hapten Mammalian Cells pANAP AnapRS Mammalian Cells Note:...intervening proteins, lipid bilayers, organelles or other cellular impediments. FRET has been used in many...endogenous biomolecules in cells nonspecifically, and they cannot cross cell membranes, so they are best... ion channels, or cell surface receptors) or with experiments on permeabilized cells. Biarsenical labeling...these compounds cross cell membranes, thus enabling FRET measurements in intact cells. The Gradia Lab has...-72 Intact Cells Cy3NTAc Poly His  44 35-63 Plasma membrane protein, permeabilized cells  Cy5NTAc ...-70 Intact Cells Cy3NTAc Poly His  63 50-90 Plasma membrane protein, permeabilized cells  Cy5NTAc ...

...Non-Dividing Cells Transient Gene Expression Low Tropism for Mouse Cells and Hematopoietic Cells Low Packaging...cargo” to target cells and organisms, and when that cargo is designed to make you sick…well, you feel it....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...Viruses as biological tools In nature, viruses infect cells and organisms (which we refer to as hosts) via viral...

...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...

...With regard to target cells or organism, some primary cells and differentiated cell types are picky when...packaging plasmids into a viral production cell line, usually HEK-293 cells. When all of these viral plasmids ...secreted into the cellular media, making viral collection fairly easy. Producer cell media can be collected... rogue HEK-293 cell). This virus-containing media can then be placed on whatever cells you would like ...just zapping your cells? That really depends on two things. First, what type of cell or organism context...viruses as well. Transduce on! Resources Resources on Addgene.org Generating Stable Cell Lines with ...process of transferring foreign nucleic acid into a cell by a viral agent. Transductions are similar to transfections...

...prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells. The... continuous cellular processes (e.g., protein synthesis), while amp only affects the cell wall synthesis...got amp? Well, amp is less stable than its counterparts and is also known to produce satellite colonies...decorative" title="Check icon" %} Satellite colonies Kanamycin Excellent No Bacteria and eukaryotes (G418...Check icon" %} Spectinomycin Excellent No Just bacteria Not SHuffle cells {% icon icon_set="fontawesome...icon" %} Zeocin Excellent No Bacteria, plants, and eukaryotes Not Tn5-containing cells {% icon icon_set..., penicillin, and is a beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces...

...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.   ...

... 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...great performance in cellular functional imaging and in protein fusions. The Gadella lab has deposited all...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...

...therapy and ex vivo cell engineering applications and is especially well-suited for lung cells via airway administration...epithelial cells, liver, CNS (limited penetration), retina, hematopoietic cells, and immune cells. B) AAV11...Sanchez, P. S., & Rodriguez-Fraticelli, A. E. (2025). Pre-existing stem cell heterogeneity dictates clonal...the acquisition of leukemic driver mutations. Cell Stem Cell, 32(4), 564–580.e6. https://doi.org/10.1016...you'll find in this post: Bright and photostable yellow fluorescent protein mGold2 Viral delivery of TnpB...plant genome editing LARRYv2 pooled libraries for cell barcoding Addgene’s own AAV6 and AAV11 packaging...connections with ATLAS Bright and photostable yellow fluorescent protein mGold2 By Mike Lacy Get your...

...PV-positive cells out of all BiPVe3-dTom cells. C) Confocal stack of a biocytin-filled cell expressing ...particles in HEK293T cells with pMD2.G (Addgene #12259) and psPAX2 (Addgene #12260). HeLa cells were infected...brightness, kinetics, and sensitivity. iGluSnFR4f is well-suited for imaging rapid synaptic events such as...Scheme for two-photon (2P) imaging and loose-seal, cell-attached recordings in primary visual cortex (V1...contains everything needed to replicate inside a host cell but with space to insert your genetic cargo of choice...promoter drives robust gene expression in mammalian cells. Flexible tagging: Choose untagged or add a versatile...stable: A puromycin resistance gene makes stable cell line creation a breeze. Modular by design: Key elements...

...lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic material...particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor and sometimes ..., 2006). AAV6 is particularly well-suited for transduction of lung cells via airway administration (Halbert...has tropism for cardiac and liver cells (Issa et al., 2023), as well as skeletal muscle, although muscle...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....

...linkage of intra- and extracellular environments. Found on nearly every cell in the human body, integrins are...array of processes, including cell adhesion, migration, and signaling, as well as immune response, tissue... that protrude from the cell membrane, physically connecting the extracellular environment and the cytoskeleton...ligand binding, mediating cell adhesion and migration, initiating intracellular signal cascades, and allowing... the extracellular matrix. During immune responses, this same conformation encourages immune 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 ...

... is technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to...its propensity to bind cellular components. Some dyes target specific organelles like mitochondria, and... 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... dye or ligand to make your selection easier, as well as highlight some recent advancements within this...collection contains many self-labeling protein fusions as well as empty backbones to clone your own protein of ...

...matters. Well, it turns out this isn’t the case for using AAV. When it comes to infecting cells, titer,...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 ...

...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 ...exposed to plant cells. The region of the Ti plasmid between the T-DNA borders (yellow) are recognized ... shown to be excellent alternative genome editing technologies especially in human cells. In contrast ... 2019) or cell-penetrating peptides (Santana et al., 2020) and plants viruses (barley yellow striate mosaic...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...

...). These cell-permeable Janelia Fluor (JF) dyes have excellent properties inside living cells, especially... fluorescent proteins. In-cell labeling strategies The majority of in-cell labeling strategies have two...and oligonucleotide labels for fixed-cell imaging, not for live-cell applications. Thus, these fluorophores..."Imaging live-cell dynamics and structure at the single-molecule level." Molecular cell 58.4 (2015): 644...sparked a renaissance in biological imaging. Suddenly, cell biologists were no longer beholden to chemists and.... Add a dash of DNA with an electrical jolt and cells become perfectly capable of synthesizing fluorophore...question: In this age of GFP and its ilk, why should cell biologists talk to chemists? One reason is something...

...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...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...sequence serving as a functional enhancer in one cell type but not in others. The same mechanism that ...

...untreated cells—but can we be certain? Do we know whether the plasmid delivered into the cells successfully...test plasmid or transfected into a separate well of your cells. Regardless, the amount of reporter protein...amount of DNA transfected into the cells and the ability of the cells to express the protein.   In analysis...because the purified plasmid, cells, and media used in each of the wells are not independent: they were...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...