We narrowed to 859 results for: ell

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity....*Pro-Tip*: The cells will become completely confluent and the media will turn yellow. Do NOT change the...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...

...including Trends in Cell Biology, Cell, Molecular Cell, Developmental Cell, and Cell Stem Cell.  She was the...inception and launch of other Cell Press journals, including Cell Metabolism, Cell Reports and Joule. In September...Deborah Sweet from Cell Press. Deborah Sweet is the Vice President of Editorial at Cell Press.  She trained...the founding Editor-in-Chief of Cell Stem Cell, a role she held for 10 years, and played a major role in...by Deborah Sweet, Vice President of Editorial at Cell Press. Almost everyone who works in a lab struggles...and making sure you do what you can to help your fellow researchers advance science overall. Communication...communication of science very seriously. That’s why at Cell Press we developed STAR Methods to improve the structure...

... brain, robust single-cell labeling is necessary to distinguish individual cells. The Brainbow-1.0 construct... May 5. PubMed. Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish... both of Harvard’s Department of Molecular and Cellular Biology, harnessed these attributes of Cre-lox... 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing in other systems, such...Brainbow highlights the trend in biology towards single-cell resolution imaging. Sanes and Lichtman endeavored...contains three fluorescent proteins: RFP (red), YFP (yellow) and M-CFP (membrane tethered cyan). Without Cre...caused partially by fluorophore photoinstability, as well as the tendency of fluorophores to aggregate in ...

...specific cell type rely on transgenic models that express recombinases like Cre or Flp in their cells of interest...entry into the cell (Dudek, AM. 2018).  If the serotype you are using transduces cells that you are not... an ATG start codon to prevent transcription in cells where the recombinase was not present (Kuhlman, ...recombination that can lead to leaky expression in cells that do not express Cre or Flp. Here at Addgene ...recombined/functional transgene that can be expressed in cells where Cre is absent. Additionally, we have experienced.../Flex vectors and leaky expression can occur in cells that don’t express Cre or Flp, what can we do about...volume needed to achieve adequate expression in your cells of interest.  The AAV serotype used can exhibit ...

...PFs need to tell and sell their stories to get jobs. PFs (and mentors) can never be too well practiced ...doing a postdoctoral fellowship, what are you signing up for and how can you do it well? Below, I briefly...exciting story I can tell with my data?  Can I make better slides that more effectively tell a story? Generally...Hughes Medical Institute. Today's postdoctoral fellows (PFs) face a number of challenges ranging from ...briefly describe some tips for a successful postdoc fellowship. 1. Reverse engineer success On Day 1 of your...preparation) A history of successful funding (fellowship grants) A project with sufficient preliminary...need to be there? A typical biomedical postdoc fellowship lasts five years. Most universities and institutes...

... cultured cells as well as more difficult conditions such as model systems and primary cells. References...when trying to screen a limited number of cells like primary cells or in vivo. Recently, the labs of David... human CRISPR libraries. Brunello: human CRISPRko sgRNA library The Brunello library consists of 77,441...criterion for probing a limited number of cells. Get the Brunello CRISPRko sgRNA library as pooled library...with both A375 (melanoma) and AT29 (colon cancer) cells and assessed the libraries performance via the depletion... viability screens. Based on these metrics the Brunello library was more effective than other popular ...genes. In the subsampling analysis of the sgRNAs, Brunello even with only a single sgRNA outperformed another...

...measure, and control molecular signals, cells, and groups of cells in order to understand their activity...Fluorescence-activated cell sorting (FACS): This is a type of flow cytometry that separates mixtures of cells into distinct...Thus, FACS can be used to separate cells expressing GFP from cells that are not. Developmental/transgenic...(GFP) has likely existed in creatures such as jellyfish for millions of years; however, it took until ... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing...consisting of 238 amino acids derived from the crystal jellyfish Aequorea victoria. It has a fluorescent emission... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...

...of interest into cells. Unlike gamma-retroviruses that can only infect dividing cells, lentiviruses can...dividing and non-diving cells while retroviruses can only infect dividing cells. When choosing the appropriate...to ensure that the proper cell line is used; several common laboratory cell lines including 293 were immortalized...can infect dividing and non-dividing cells.  Addgene has an extensive collection of lentiviral plasmids...transfecting four different plasmids into the producer cells (two packaging plasmids, an envelope plasmid, and...pol, and env), the isoforms of these proteins, as well as the viral long terminal repeats (LTRs) are different...appropriate gene delivery system, the specific target cell or tissue needs to be considered. Q2: Can I use...

... Oct 7, 2020 by Gabrielle Clouse. This post was contributed by guest bloggers Marcelle Tuttle and Alex...differentiate induced pluripotent stem cells (Chavez et al., 2015) into neuronal cells by activating genes that cause...drug targets by finding genes that cause cell death in cancer cell lines upon activation (Gilbert et al.,...reasonably well.  Best of luck on your experiments!  Many thanks to our guest bloggers Marcelle Tuttle ...Repression and Activation. Cell 159:647–661 . https://doi.org/10.1016/j.cell.2014.09.029 Konermann S, Brigham...and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/10.1016/j.cell.2014.09.039 Wong N, Liu W,...activation allows for exploratory-based research as well. For example, CRISPRa can be used to find potential...

...homodimerize in cells. To understand how well fluorescent proteins approach this ideal situation, cellular assays... 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...a non-aggregating fluorescent protein. In the jellyfish Aequorea victoria, AvGFP forms a homodimer. In...tolerated in some applications (e.g. labeling of cells or tracking promotor activity), but it is problematic... affinity and has been used to demonstrate that yellow fluorescent protein homodimerization is characterized...fluorescent proteins interact or oligomerize in cells? This question is largely unanswered. Ideally, fluorescent...

... origin as well as a selectable marker and a desired genetic payload (ex: GFP). Thus, cells can be both...bee gut microbiome species such as Snodgrassella alvi and Gilliamella apicoli. The plasmids could also ...strategy for bacteria to count cell divisions. The technique, called distributed cell division counting (DCDC...one bright particle that segregates into one cell during cell division. To achieve the agglomeration of ...promoter. Cells are first induced with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are... new particles. With subsequent cell divisions, the fraction of cells containing the particle exponentially...plasmids to both Gram-positive and Gram-negative cells.  The Wang lab further developed a modular suite...

...successfully infected/expressing cells by flow cytometry. The isolated cells were injected into mice and imaged...including monitoring multiple cellular events such as the dynamics of subcellular structures and gene expression...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors...acceptor. NLuc is an excellent BRET donor due to its bright luminescence. Intracellular Calcium Sensor Yang...to changes in intracellular calcium and is useful for direct imaging of events in cells and brain explants...could “clone by phone”. One of my professors was excellent at phone cloning with labs around the world and...luciferase by Promega’s Advanced Technologies group as well as evolution of the coelenterzine substrate to the...

...active wavelengths to reach zooxanthellae, the coral’s symbiotic dinoflagellates that are photosynthetic ... them into your cells, you can simply look for blue colonies to determine which cells have taken up the... that the cellular fitness cost varied greatly between chromoproteins. In some cases, cells containing... mScarlet as the donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you... is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful corals...described an unusual blue-colored protein from the jellyfish Cassiopea xamachana (Blanquet and Phelan, 1987...cloning experiment worked just by looking at the cells on the plate. For example, if your plasmid contains...