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We narrowed to 1,029 results for: ELL

Showing: 281 - 320 of 1029 results

  1. Addgene's Cancer Collection Pages Connect the Research Community

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    Blog Post
    ...molecular level, at the cellular level there is a universal theme - uncontrolled cellular proliferation. We ...cancerous cells Luckily, there are several safety mechanisms in place to protect us from rogue cells when ...process of programmed cell death, or apoptosis, is one such mechanism. When a cell or its genetic material... directly inhibit the ability of a cell to undergo programmed cell death while simultaneously stimulating...eliminate every cancerous cell. Even so, to pose the greatest threat, cancerous cells still need to successfully...universally-dividing behavior of cancer cells, attacking the cellular machinery necessary for division. This...proteins that, when expressed, normally encourage cell survival, growth and proliferation, while tumor ...

  2. Genome-wide Screening Using CRISPR

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    Blog Post
    ...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...

  3. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    Blog Post
    ...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 ...

  4. Viral Vectors 101: Voltage Indicators

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    ...Cellular communication and function depend on potential energy. Cellular activities like action potentials...Indicators As previously mentioned, cellular voltage is measured at the cell membrane, and this voltage is ...endocrine cells. It follows that the specific targeting of these voltage indicators needs to be cell type ...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...

  5. The Fluorescent Vegetables in Aptamer Soup

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    ...challenging to track intracellularly, but aptamers permit the study of intracellular RNA dynamics. Aptamers...RNA aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged... been used to investigate intracellular viral DNA dynamics in neuronal cells (Nilaratanakul et al., 2017...be used to simultaneously target cellular RNA to specific intracellular locations using Cas9 and track ...PMC4244833. Kellenberger, Colleen A., et al. "RNA-based fluorescent biosensors for live cell imaging of...and later introducing the small molecule into the cell. The fluorescence that is generated allows for qualitative...). DFHBI is membrane permeable and non toxic to cells. Because it rapidly photobleaches, DFHBI can only...

  6. An Introduction to Adenovirus

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    Blog Post
    ... and hexon), as well as other intracellularly-exposed proteins and downstream cellular events, all recruit... AdVs is dependent on modulation of the host cell’s cell cycle; inhibition of the innate immune system... in the producer cells where there is E1 and E3 gene complementation. In target cells that are infected...death of adenoid cells in vitro (Rowe et al., 1953). While overt death of the targeted cells is less likely... & Verma, I. (1998). Cellular immune response to adenoviral vector infected cells does not require de ...infection for efficient cell lysis and release of adenovirus from infected cells. J Virol, 70(4), 2296.... some unprecedented combination of the three (Servellita et al., 2023). Normally when children have acute...

  7. Hot Plasmids - November 2023

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    Blog Post
    ... between living cells by TransitID. Cell, 186(15), 3307–3324.e30. doi: 10.1016/j.cell.2023.05.044. PMID... efficient prime editors. Cell, 186(18), 3983–4002.e26. doi: 10.1016/j.cell.2023.07.039. PMID: 37657419...lentivirus and MMLV gag, to specific cell types. It can also make CAR-T cell therapy more accessible by simplifying...Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching an old...called DIRECTED (Delivery to Intended REcipient Cells Through Envelope Design). This modular approach ...viral surface, expanding the range of targetable cell types. DIRECTED is compatible with fusogens from...process and could even enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can ...

  8. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...of proteins, organelles, and cells. Similar to FPs, PA-FPs allow for non-invasive labelling, but in contrast...Tetrameric PA-FPS are better for whole cell imaging and studying organelle trafficking. Monomeric PA-FPs are...localization, turnover, and trafficking, as well as organelle trafficking and dynamics (fission, fusion,...offer scientists a simple yet powerful way to tag cellular proteins and investigate protein localization,...continuously synthesized and degraded within the cell, so at any given time, an FP-chimeric protein may... reported was PA-GFP. PA-GFP was derived from Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating Threonine...Since emission of green light is more phototoxic to cells, most scientists working on in vivo imaging models...

  9. Viral Vectors 101: Biosensors

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    ...the caffeine of the cell!)—ATP. There are biosensors for intracellular, extracellular, and even the ADP:... as proxies for a cellular process or directly measure a cellular process. Cellular phenomena such as ... measure real-time cellular reactions to stimuli and as well as changes in cellular dynamics during so-called...type of cells the viral particles infect. This can direct your biosensor to specific tissues or cell (sub...biosensor is the thing that you care about within your cellular or animal system. This ‘thing’ could be a neurotransmitter...blog we will focus on those which can be applied in cell culture and animal models primarily. Fun fact: Almost...These ion sensors can be targeted to different cellular locations or tissues depending on the biology ...

  10. Five Popular Model Organisms

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    ... properties and processes with our own cells. Like human cells, yeast DNA is packaged into chromosomes...differentiation of β-cells in the the developing zebrafish pancreas. They first tagged these cells using plasmids...By Alyssa Cecchetelli and Lukas Morgan Model organisms are vital tools used by researchers around the...recapitulate the complex interactions between cancer cells, therapeutic drugs, and the rest of the body than...than studies in other model organisms or cell culture. For example, Scott Lowe’s lab utilized mice to study...to identify a set of mutations that made cancer cells more resistant to chemotherapy. You can find these...to optimally control gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces...

  11. Fluorescent Proteins 101: When GFP lets you down

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    Blog Post
    ...fluorescent in acidic organelles (Figure 1), showing that its acid tolerance is maintained in cells.   GFP is big...proteins to study mRNA trafficking in living cells." Methods in cell biology 85 (2008): 273-292. PubMed PMID...genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed... fluorescent protein in almost all organisms and cell types, (ii) no need to add a co-factor, (iii) easy...toleration in fusion proteins. Since GFP is such a well-validated, all-round good performing probe, it is...introducing two mutations (F64L, S65T) in the original jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant... the bathing solution and require uptake by the cells. Which of these strategies works best for a specific...

  12. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using the U-containing strand..., which may lead to cell death rather than NHEJ repair, thus removing these cells from the edited population... starting point, Gaudelli evolved the base editor into 40 new ABE8 variants (Gaudelli et al., 2020). Compared...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual base...editing than ABE7.10 Gaudelli et al., 2020 ABE8e   Efficient editing in primary cells Zhang et al., 2020...., Peraro, L., Young, L., Lee, S.-J., Ciaramella, G., & Gaudelli, N. M. (2023). Improved cytosine base...relatively low frequency, especially in nondividing cells. Scientists everywhere wistfully imagined being ...

  13. Antibodies 101: Producing Recombinant Antibodies

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    ...and yeast cells are cost-effective and faster-growing than mammalian cells, but mammalian cells can perform...your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture...production system, cell lines typically used include Human Embryonic Kidney (HEK) cells and Chinese Hamster...An antibody that performs well in immunohistochemistry may not perform well in a western blot. This can...recombinant antibodies with some molecular biology and cell culture experience. Let’s go over the basics of ... place in bacterial, yeast, plant, or mammalian cells. Each of these systems has their advantages and ...post-translational modifications that bacterial and most yeast cells cannot. Your choice of production system may depend...

  14. Supporting Rare Disease Research with the 2020 BeHEARD Awards

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    Blog Post
    ... trigger the activation of dendritic cells, macrophages, T cells, and the recruitment of neutrophils. ... says. In induced pluripotent stem cells derived endothelial cells from women who are missing an entire...detect molecules outside of the cell and translate those signals into cellular responses. For example, GPCRs...estimated that CTD is the second cause of male intellectual disability after Fragile X. There is no cure...maintaining the nutrient and ionic balance of the cells,” says Dr Sangeetha Iyer, ACD’s Scientific Advisor...that solute channels typically expressed on other cell types can compensate for or bypass the dysfunction...collection to perform a functional screen in mammalian cells to identify any solute channels that can compensate...

  15. All in a Twist: dsRNA

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    Blog Post
    ...mammalian cells. Here, we will cover how these structures are generated, why it’s important for cells to sense...that doesn’t involve cellular dysregulation) is the mitochondria. These organelles have their own circular...., Hur, S. Cellular origins of dsRNA, their recognition and consequences. Nat Rev Mol Cell Biol, 23: 286...are the words that describe RNA! Double stranded? Well, that’s just for DNA – right? RNA performs almost...almost all of its biological functions in our cells in the single strand form, but double stranded RNA and...with bidirectional transcription. In an uninfected cell, dsRNA rarely circulates, but upon viral infection...clinical therapy to target pathogenic proteins, as well as in the lab to deplete expression of proteins ...

  16. Plasmids 101: Plasmid Incompatibility

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    Blog Post
    ...copy number in the cell is high, but allows replication when copy number in the cell falls too low (Novcik...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...eukaryotic cells). The partition NTPase is recruited and “walks” each plasmid to a separate daughter cell. When...to an identification issue: the CBPs in the cell can’t tell the difference between plasmids if they have... daughter cell, instead of partitioning each type of plasmid into a separate daughter cells (Figure 2)...from the cell with the same probability, or asymmetrical, where one plasmid lost from the cell at a higher...different plasmids to be maintained in one bacterial cell. In this Plasmids 101 post, we’ll cover why this...

  17. Predicting Adverse Reactions to Monoclonal Antibody Drugs

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    ...epitopes likely to be presented by T cells to elicit an immune response. T cell epitopes are found in either ...predict T cell epitopes focus on characterizing those epitopes as CD8 or CD4 epitopes, as well as their...that take place within an antigen-presenting cell like a T cell. It is also not yet possible for these algorithms...traditional 2D assays with immune cell lines or peripheral blood mononuclear cells (PBMC) in 3D models of the...CD4+ or CD8+ T-cells or monocytes. PBMCs can be further differentiated into dentritic cells or macrophages...skin and subcutaneous models (Groell et.al, 2018). Treating these cells with the therapeutic antibody ..., that are commonly used for the prediction of T cell epitopes based on the amino acid sequence of the...

  18. Fluorescent Tagging of Endogenous Genes with SapTrap

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    ...associated with cellular organelles, scientists have been able to study many cellular processes, including...and function of cellular systems. FPs have been used to track protein localization, cell structure, intracellular...to fluorescently tag markers of cellular structures in mammalian cells. These plasmids use fluorescent...Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors.” Cell Rep.15(1) (2016...trafficking. Although FPs have enabled discovery of many cellular mechanisms, there are some limitations to working... ultimately misinterpretation of the protein’s cellular role. One way to avoid the pitfalls of overexpressed...synthetic DNA for the desired gRNA target sequence, as well as the 5’ and 3’ homology arm repair template (Fig...

  19. Fluorescent Proteins 101: Fluorescent Protein Timers

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    Blog Post
    ...to monitor cell, organelle, and protein localization. For instance, you might dye your cells and look at...either in whole cells or as a pool of fusion proteins localized to a specific region of the cell. For example...the relationship between the age of a cell, protein, or cellular structure and a particular biological...trafficking to a subcellular location, start of gene expression, development of a cell structure, etc)....biology, FP timers might be able to tell researchers whether old or new cells are better at producing a compound...microscope, fractionate samples to isolate particular organelles and their contents, or perform in situ hybridization...timers, researchers can look at a single image of a cell and understand how protein localization changes ...

  20. 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... not well understood, researchers have been using AAV vectors for gene editing in mammalian cells since...transient transfection of 293T cells with helpers, and used to infect recipient cells. After G418 selection, ...since 1998. In that year, Russell and Hirata from the University of Washington reported a surprising finding...used to routinely deliver sequences of interest to cells with efficiencies of 1-40% after G418 selection ...composed of single stranded DNA, which is particularly well suited for strand invasion, a key step in the homologous...the p53 and PTEN tumor suppressor genes in human cells (Kim et al 2008). They have also used AAV gene editing...

  21. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    ...packaging cell line. γ-Retroviruses can transduce target cells, but the transduced target cell does not...using helper-free packaging cell lines This method utilizes a packaging cell line that stably expresses... packaging cell line. For example, Phoenix, a second generation γ-retrovirus packaging cell line developed...and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for the infection of mammalian cells.) Using...transduce both dividing and non-dividing cells because once in the host cell cytoplasm, the viral components are...for efficient gene delivery to mammalian cells using 293T cell-based systems. Curr Protoc Immunol. 2001...γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted the development of multiple...

  22. Lentiviral Vector Uses and Overview

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    ... of cell types, so they’re ideal for work with primary cells or other difficult-to-transfect cells. Lentiviral...dividing cells. Another type of vector, based on adenovirus, could infect non-dividing cells, but without...turned to the lentivirus HIV-1, well-known to infect non-dividing cells. The first lentiviral vector system...ability to transduce both dividing and non-dividing cells. Lentiviral vectors are the vector of choice for...viral envelope to dictate the tropism, or types of cells the vector could infect. Finally, the transfer plasmid.... After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles...adeno-associated viral vectors can also target non-dividing cells, lentiviral vectors can carry much more genetic ...

  23. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

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    ...lines expressing GFP in specific cell types to do more than just label cells. If GFP could be co-opted to ...reporter output was observed in their 293 cell culture system. In cells cotransfected with GFP and the GBPs,...expression was induced in ~76% of GFP+ cells, and 100% of DsRed+ cells also expressed GFP. Using AAV constructs...cortex, cerebellum, and spinal cord. Cre-DOG is also suitable for optogenetic studies, and since cells infected...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed ...wanted to express a variety of proteins in single cell types in the mouse, but knew it would take years...could then selectively manipulate only GFP-labeled cells. Once Tang and Cepko found a description of GFP-...

  24. Fluorescent Biosensors for Measuring Autophagic Flux

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    ...currently available from Addgene. Rosella autophagy biosensor Rosella is a dual-emission autophagy biosensor... autophagy, Rosella is targeted to the vacuole in yeast or the lysosome in mammalian cells. This decrease... and cytoplasmic Rosella. When fused with a mitochondria-targeting sequence, Rosella can be used to track...al. "Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in yeast...process by which cytoplasmic material, including organelles, are targeted to lysosomes for degradation. Autophagy...provides limited information about what materials or organelles are being targeted for autophagy. The development...). In the absence of autophagy, the untargeted Rosella remains in the neutral pH cytoplasm where it emits...

  25. 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...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...such as Na+, K+, and Ca++ to passively enter the cell with the concentration gradient to depolarize it...

  26. Antibodies 101: Designing Your First Flow Panel

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    ...and inside the cells and are thus typically used for distinguishing live from dead cells. The combination...the chart tells us that PE and PE-Cy7 are "very bright" when they are excited by the yellow/green laser...say I am culturing HEK 293T cells in vitro. I transduced the cultured cells with a virus that encodes a...panel. Besides some background cell death constantly happening in cell culture, the transduction process... to some cell death. As a result, there is a high abundance of positively stained dead cells present and...When analyzing your cells using flow cytometry, you are typically measuring the presence or absence of...certain markers on the surface or the inside of your cells. While proteins themselves can emit intrinsic fluorescence...

  27. Hot Plasmids and Viral Preps - January 2021

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    ...fluorescence-activated cell sorting (FACS) helps researchers sift through thousands of cells to identify cells exhibiting... CRISPR-Casɸ is active in vitro as well as in human and plant cells. Casɸ from Biggiephage can ...target single cells. Now, Francois St-Pierre’s lab developed a new form of FACS-based cell screening, Single-cell...individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures.  HeLa cells expressed...was used to image over 3 million cell types expressing variants of Yellow Fluorescent Protein (YFP). The...Cre for use in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items... conformational changes in a protein.    Cells expressing an RFP reporter plasmid for recombinase...

  28. CRISPR in the Clinic

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    ...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (2013). ...patient’s own cells that have been modified ex vivo, previously edited healthy donor cells can be delivered... editing these cells to avoid a negative immune response (e.g., graft vs. host) as well as the introduction...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (2013). Gerald Schwank et al. ...on editing animal models of disease and cultured cells. For example, in 2013, Jinsong Li and colleagues...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis ...delivery directly to the patient. Patient-derived cells can alternatively be harvested, edited ex vivo, ...

  29. Creating accessible biology activities in schools with BioBits

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    ...on freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free extracts Typically...organism. Furthermore, cell free systems can be freeze-dried into pellets (FD-CF pellets), leaving them shelf-stable... into a host cell (through a process called transformation), essentially giving the cell all the instructions...to dispose of the live cells. The alternative is to express the protein in a cell-free manner. This is ... about operating the rest of the car as well. Similarly, cell free expression systems contain all the ...first molecular biology lab in college that I really fell in love with it. Something about being able to actually...biology, especially those at the molecular and cellular level. The reason for this is cost and complexity...

  30. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

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    ...targeted One Many Sample type(s) Cell lines Cell lines, primary cells Table 1: Key differences between... of cells (~5x107). Together, these two requirements prevent the use of CAPTURE with primary cells or ...need to create stable cell lines and allows CAPTURE 2.0 to be used with primary cells. Find plasmids for...that occur during development and cellular differentiation. Using a cell line model, the Xu Lab tracked ...by Biotinylated dCas9. Cell 170:1028–1043.e19 . https://doi.org/10.1016/j.cell.2017.08.003 Liu, X., Chen...CAPTURE method requires the creation of a stable cell line that co-expresses three components: dCas9 ...drawback of the original CAPTURE method is that a new cell line has to be created for each genomic loci of ...

  31. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    ... specific cell-type promoter. Once injected into the brain, the viruses infect all the cells with an inactive...the brain remained elusive, not from a lack of intellectual curiosity on the part of scientists but, rather...energy conversion or the induction of intra- or intercellular signaling pathways (Ernst et al., 2014). Microbial...proteins control the flow of electric charges across cell membranes and maintain membrane potential in response...once a certain number of positive ions crosses the cell membrane, a threshold is reached and the neuron ...selective expression of opsin genes in targeted cells makes it possible to control neuronal activity with...that, in turn, trigger neural excitation, whereas yellow light activates halorhodopsins to silence neuronal...

  32. Sequencing Options for CRISPR Genotyping

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    ...Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with cell-based method ...Requires large cell population, senstive to time since cell fixing HTGTS Not reported Cell-based Identifies...method IDLV 1% Cell-based Programmable, can detect DSBs in live cells Not as senstive as other unbiased methods... use single cell clones for in vitro CRISPR experiments. The likelihood that a single cell clone derived...and cannot be applied to heterogenous populations (Bell et al. 2014). Further, Sanger sequencing has a lower...product sequence. Targeted amplicon sequencing doesn’t tell the whole story Figure 1: Schematic representations...picture of the impact of CRISPR editing in the model cell line or organism. Several options exist for unbiased...

  33. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

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    ...introduced the reporter into the cells, you’re ready to edit. These cells are then transfected with several... up, all successfully transfected cells will express RFP while cells containing the reporter gene that...cut and repaired by NHEJ as well. The ratio of RFP to GFP expressing cells provides a real-time read out...using the RFP:GFP ratio, edited cells can be sorted from unedited cells. In an experiment that targeted...Activity. Frontiers in Cell and Developmental Biology 6: . https://doi.org/10.3389/fcell.2018.00054 Sander...CRISPR experiment was successful in your organism or cell type? You can use DNA sequencing or other molecular...histone)-GFP reporter into the genome of mammalian cells.  Figure 1: The SRIRACCHA reporter construct...

  34. Viral Vectors 101: Types of viruses

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    ...dividing cells only, so is not suitable for infection of quiescent cells or infrequently cycling cells but ...non-dividing cells, making it conducive to targeting all cell types. Altogether, AAV has many excellent features...DNA copies of their genome upon infecting a host cell. This viral DNA (also called proviral DNA) is then...host. So why use a gamma-retrovirus in the lab? Well, they are essentially evolutionarily optimized gene...and permanently integrate a genetic cargo into a cell. These viruses have a cargo capacity of ~ 8 kb, ...needs, and can transduce (infect) a wide range of cell types with a high efficiency. However, gamma-retrovirus...but is fine for experiments with immortalized cell line cultures. Lentivirus Lentivirus is a type of retrovirus...

  35. Antibodies 101: Fc Effector Functions

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    ...animation.)  Many types of immune cells, from mast cells to B cells, express Fc receptors that bind to...) antibody-dependent cellular cytotoxicity (ADCC) c) antibody-dependent cellular phagocytosis (ADCP). ...opened in the pathogen’s cell membrane (Dunkelberger & Song, 2010). As cells cannot survive when holes...die.  Antibody dependent cellular cytotoxicity (ADCC) Antibody dependent cellular cytotoxicity (ADCC) is...(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...

  36. Fluorescent Proteins 101: Fluorescent Biosensors

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    ...looking at an intracellular or extracellular molecule? By fusing a biosensor to an organelle-specific targeting... fluorescence across a population of cells (i.e. in a plate well). Use to track a process (e.g. enzyme...Localization within cells Fluorescence microscopy Visualize components of individual cells Separation/identification...fluorescently-labeled components Fluorescence-activated cell sorting (for cells); fluorescent HPLC (proteins or other molecules...) Isolate or quantify cells, proteins or other molecules that have been labelled with your fluorescent...signal. Blue flame GECO biosensors from Robert Campbell’s lab display calcium-dependent increases in fluorescence...by Olivier Pertz’s lab. Other biosensors track cellular characteristics like pH, redox state, and voltage...

  37. New Tool for Lineage Tracing: The ClonTracer Library

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    ...resistant cancer cells exist prior to drug treatment. They infected a non-small cell lung cancer cell line containing...same parent cells that represent the preexisting resistant subpopulation 2. Resistant cells among the separate... target cell population at low multiplicity of infection (MOI) such that each individual cell receives...researchers to individually label millions of mammalian cells through lentiviral infection and to monitor their...quantify the take rate of mouse xenografts after cell implantation. Their goal was to figure out how to...ClonTracer library to determine whether or not cancer cell populations resistant to drug treatments arise during...They hypothesized that, if they labeled the parent cell population using the library, propagated it, separated...

  38. Hot Plasmids - August 2020

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    Blog Post
    ...structure or protein is located within a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing...for labeling specific structures within cells. HeLa cells were transfected with plasmids encoding...affinity), as well as GRAB_NEmut as a negative control. Find the sensors at Addgene! New yellow fluorescence...protein evolution Fluorescent tools for mammalian cells Nanobody purification CRISPRi and CRISPRa system...selection in the cytosol (instead of on the yeast extracellular surface) which enables tracking of protease ...mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image from Mateo...labeling structures and compartments in mammalian cells By Angela Abitua Genetically encoded fluorescent...

  39. Viral Vectors 101: Preparing Pooled Libraries

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    ...transduction of cells, resulting in cell population mutated by the viral pooled library. These cells can then...transduce your cells.  Titering After generating your lentivirus, you’ll want to infect your cells with the ...of a cell being infected by only one viral particle, but it also increases the chances that a cell doesn...adequately represented in the cell population, you’ll need to infect many more cells than the number of plasmids...appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus. Purification of...small aliquot is not enough to transfect producer cells, so your first step will be to amplify the library...individual plasmids knows, some plasmids amplify well and others don’t. The size and composition of your...

  40. Hot Plasmids and Viral Preps - September 2021

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    ...improved cell-free system to examine biosynthetic pathways A new tool for enriching ABE-targeted cells The ...Improved cell-free transcription/translation system to examine biosynthetic pathways  By Gabrielle Clouse...interest, and cell extract.    Graphical Abstract: Schematic representation of cell-free system,...successfully reconstituted in this cell-free system. In summary, this cell free system allows for a relatively...   New tool for enrichment of ABE-targeted cells By Ellie Adams Single base modifications through ABE...multiplexed and can be used in human pluripotent stem cells, a cell type that has been difficult to modify in the...Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems) have been...
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