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We narrowed to 819 results for: cell

Showing: 121 - 140 of 819 results

  1. Adenoviral Vector Production and Troubleshooting

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    Blog Post
    ...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity...more HEK293 cells and produce more rAdV to reach higher titers. Successfully infected cells will become...large transgenes, transduce quiescent and dividing cells, and do not integrate into the host’s genome. The...administration, which results in the death of transduced cells and loss of transgene expression (Interestingly,...gene delivery: Potential applications for gene and cell-based therapies in the new era of personalized medicine...can be generated if the E1 gene from the packaging cell line is transferred into the AdV by recombination...stock by doing a plaque formation assay on A549 cells . Alternatively, antibody-based assays are commercially...

  2. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...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 (...

  3. Special Delivery: Fluorophore Targeting for FRET Studies

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    Blog Post
    ...Mammalian Cells pCMV-DnpK dinitrophenyl hapten Mammalian Cells pANAP AnapRS Mammalian Cells Note:...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 ...-78 Intact Cells Cy3NTAc Poly His  65 52-94 Plasma membrane protein, permeabilized cells  Cy5NTAc ...

  4. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

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    ...Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific expression of molecular tools...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...with AAV: 1. Getting your viral vector into your cell of interest: AAV tropism and route of administration...transduction, the viral capsid proteins bind to the target cell’s surface proteins and this interaction determines...efficiency of the AAV serotype used for a specific cell type. Find some transduction characteristics of ...by restricting expression to genetically defined cell types. Specific transgene expression can be also...different approaches can increase the specificity of cell targeting even further. For example, injecting AAVs...

  5. Immunology Research Plasmids and Resources

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    Collection
    ...foreign antigens to adaptive immune cells, such as T cells and B cells. About a week following the initial...presenting cells. B cells in secondary lymphoid organs recognize specific antigens through the B cell receptor...have been described. NK Cell Signaling Natural killer (NK) cells can kill cells that have been infected... within an infected cell membrane, triggering cell death. TCR Signaling The T cell receptor (TCR) engages...through effects on numerous immune cell types including T cells and B cells. It also impacts the development...function of innate immune cells including NK cells, macrophages, dendritic cells, and granulocytes. TLR ...

  6. Antibodies 101: Fc Effector Functions

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    Blog Post
    ...animation.)  Many types of immune cells, from mast cells to B cells, express Fc receptors that bind to...opened in the pathogen’s cell membrane (Dunkelberger & Song, 2010). As cells cannot survive when holes...(NK) cells expressing Fcγ receptors binds to IgG and release cytotoxins that kill the target cell. If ...response, by binding to Fc receptors on other immune cells. (psst! Need a quick review of the different immune...pathogen and physically prevent it from entering a cell, a non-Fc-dependent effector function known as neutralization...pathogen and mark it as a target for other immune cells, in a process known as opsonization. In antibody-dependent...shaping the immune response by driving particular cells or responses. And there are still antibody isotypes...

  7. Hot Plasmids: Summer 2025

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    Blog Post
    ...epithelial cells, liver, CNS (limited penetration), retina, hematopoietic cells, and immune cells. B) AAV11...the acquisition of leukemic driver mutations. Cell Stem Cell, 32(4), 564–580.e6. https://doi.org/10.1016...therapy and ex vivo cell engineering applications and is especially well-suited for lung cells via airway administration...Sanchez, P. S., & Rodriguez-Fraticelli, A. E. (2025). Pre-existing stem cell heterogeneity dictates clonal...plant genome editing LARRYv2 pooled libraries for cell barcoding Addgene’s own AAV6 and AAV11 packaging...filaments with mGold2. Top: TIRF-SIM imaging of COS-7 cells expressing LifeAct fused to mVenus, mGold2s, or ...many barcoding libraries are available for tracking cell lineage, different applications require careful ...

  8. Plan Your Experiment

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    Collection
    ...difficult-to-transfect cells. You can direct expression to specific cell types by using cell-specific promoters...human embryonic kidney 293 (HEK293) cells. For more difficult cell types, you may need to look into other...are good choices for more difficult cell types like primary cells. For CRISPR delivery, lentiviral and...deliver to target cells through microinjection or electroporation. The target cells translate the mRNA...complexes directly into your target cells. In this case, your target cells are not transcribing DNA to mRNA...possible genotypes within the resulting cell population. Some cells may remain wild type due to either (1...modifications . Cell Population The first step of validation is determining the type of cell population you...

  9. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ... quantified for individual cells, and the number of 'normal' looking cells can be counted. This assay ...living cells. Strikingly, the OSER assay reveals that tagRFP does not act as a true monomer in cells (Constantini...homodimerize in cells. To understand how well fluorescent proteins approach this ideal situation, cellular assays...tolerated in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic...fluorescent proteins interact or oligomerize in cells? This question is largely unanswered. Ideally, fluorescent...proteins could properly form microtubule networks in cells. Other efforts tested whether FP fusions to connexin...present in EGFP. Constantini et al, (2012) developed a cell-based assay that does a better job of detecting ...

  10. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

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    ...for neurons. Non-neuronal cells do this too. It's just that for non-neural cells, ion-based communication...between many cells (through gap junction proteins which connect the cytoplasm of two cells) can also affect...more depolarized (positive) the cell is, the more embryonic, stem cell-like, or cancerous it is (Levin...happens at a slightly slower scale.  All living cells with a membrane possess an electrical state. This...charge between the inside and the outside of the cell. This imbalance is maintained by ion channels and...proteins cx43 GJB1 GJB2 Bioelectric states can alter cell types and patterning There is mounting evidence ...more polarized (negative) the action potential of a cell, the more differentiated and quiescent it is, and...

  11. Antibodies 101: Isotypes

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    Blog Post
    ...the surface of basophils and mast cells, two types of white blood cells involved in allergic reactions. ... is important because it determines what immune cells and molecules are recruited by the antibody to help...each of these functions: Recruitment of immune cells that express receptors that recognize the Fc portion...special receptors that transport antibodies through cells and into different body compartments, such as into... during a primary immune response because all B cells begin by expressing IgM. IgM has a pentamer structure...coexpressed with IgM on the surface of most immature B cells, its function is unknown. It has a monomer structure...Isotype switching Isotype switching occurs when a B cell changes which heavy chain constant domain it pairs...

  12. Mapping the 4D nucleome with CRISPR/Cas9

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    Blog Post
    ...be applicable to studies of cell cycle progression, epigenetics and cellular reactions to external stimuli...Weissman JS, Qi LS, Huang B. Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001. Pubmed. Find...LS, Weissman JS, Vale RD. Cell. 2014 Oct 23;159(3):635-46. doi: 10.1016/j.cell.2014.09.039. Epub 2014 Oct...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...

  13. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    Blog Post
    ...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...

  14. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...organs, cells comprise compartments and structures, called organelles. Take a sneak peak inside a cell with...proteins, however, in this case the cell needs to be fixed and the cell membrane permeabilized in order for...Cooper GM. 2000. The Cell: A Molecular Approach. 2nd Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory...involved in intracellular transport pathways, it is possible to map the internal organization of a cell. This...for labeling your subcellular structure of interest in the Allen Institute for Cell Science Plasmid Collection...with the images from the Allen Cell Explorer (1). When studying the function of a protein or its role in...biochemical methods thus removing the context of the cell. However, much knowledge about functionality can...

  15. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...endocrine cells. It follows that the specific targeting of these voltage indicators needs to be cell type ...Indicators As previously mentioned, cellular voltage is measured at the cell membrane, and this voltage is ...electrical events! Voltage indicators are voltmeters for cells! In this blog we will review how these indicators...related to the electrical impulse of the cardiac cells. To study this phenomenon you will likely want to...the voltage change across the membrane of cardiac cells. Similarly, neuronal signaling is carried out by... electrical charges propagated through multiple cells to do things like translate perceived signals to...Understanding neuronal pathways and the function of certain cells depends on the ability to monitor their signals ...

  16. Hot Plasmids: Spring 2025

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    Blog Post
    ...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...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...step-by-step protocols for viral generation and stable cell line creation (Figure 2). Looking for more? Explore...

  17. Adeno-associated Viruses (AAVs) for Genome Editing

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    ...K562 leukemia cell line and ~8 and 9% efficiencies in T-cells and hematopoietic stem cells respectively...transient transfection of 293T cells with helpers, and used to infect recipient cells. After G418 selection, ...using AAV vectors for gene editing in mammalian cells since 1998. In that year, Russell and Hirata from...used to routinely deliver sequences of interest to cells with efficiencies of 1-40% after G418 selection ...the p53 and PTEN tumor suppressor genes in human cells (Kim et al 2008). They have also used AAV gene editing...endogenous allele of the STAG2 tumor suppressor in human cells (Kim et al, 2016). The Waldman lab has designed ...modifying multiple genes simultaneously in the same cell, are more appropriate for a CRISPR-based approach...

  18. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

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    ...incorporated into the edited genome. In mammalian cells such as stem cells, HDR is relatively rare and DSBs are predominantly...them into somatic cells affected by the disease and study what’s wrong with the cells. However, our excitement...sequence change the cell must undergo homology-directed repair (HDR), a distinct cellular DNA repair pathway...sequence-specific changes in human induced pluripotent stem cells (iPSCs). The CRISPR/Cas9 com­plex is very efficient... stranded breaks (DSBs) into genomic DNA in many cell types and often results in biallelic modifications... After genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we...excitement turned into frustration when we sequenced our cells and found how inefficiently our edits were being...

  19. 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...

  20. Mouse Modeling, Part 1: Genetically Engineered Mice

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    Blog Post
    ...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...
Showing: 121 - 140 of 819 results