...repair protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag ...Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag Control for cell surface... sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells To learn...visualization of proteins inside cells. Depositor... Depositor Collections New England Biolabs Cell-Imaging Plasmid Collection...Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental Information ...visualization of proteins inside living or fixed cells. The plasmids associated with this technology are...

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

...varying across cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher... of your cells. Enrich S-phase cells HR operates predominantly in S and G2 phase of the cell cycle, when...HDR events, ensure cells are actively cycling through the cell cycle stages. If cells are overly confluent...way to make a knock-in cell line is to utilize the built-in repair pathway cells already have for repairing...than quiescent cells. It is important to know that HR is often mutated in cancer cells, specifically in...keeping an eye on those cells! To take this one step further, you can inhibit cell cycle regulatory proteins...negative outcomes on viability if cells are arrested too long. Prolonged cell cycle arrest can lead to apoptosis...

...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... in Human Pluripotent Stem Cells: Choosing the Correct Path.,” J. stem cell Regen. Biol., vol. 1, no. ...cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,” Proc. Natl. ...

...on or in these cells: antibodies can bind to proteins on the cell surface, or the cell membrane can be...fusing B cells that have been extracted from animals injected with protein antigen to myeloma cells. The ... measurement of protein expression in cell lysates, whole cells, or tissue samples. Many experiments that...heterogeneous mixture - for example, cell lysate or media from cultured cells. ELISA can be used to quantitatively...same cell simultaneously as well as compare between samples. Learn more about flow cytometry. Cell and ... (immunohistochemistry) or cells (immunocytochemistry). This could be cells or tissue slices mounted to...an animal, to directly label circulating cells or other cells that are accessible from the bloodstream...

...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...communication of science very seriously. That’s why at Cell Press we developed STAR Methods to improve the structure...trained originally in cell biology, completing her PhD at the MRC LMB in Cambridge UK and then a postdoc...

...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...

...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...

...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...

...). 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...

...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 ...

...enzyme-mediated proximity cell labeling strategy to detect cell-cell interactions in vivo (Ge et al., 2019...about RapID! EXCELL: Proximity labeling for the detection of cell-cell interactions EXCELL is an enzyme-mediated...et al. "Enzyme-Mediated Intercellular Proximity Labeling for Detecting Cell–Cell Interactions." Journal...a bait protein fused to BirA* in cells and then incubate those cells with exogenous biotin. After a few...-protein interactions, and even more recently cell-cell interactions. If you’re looking to identify interactions...proximal and interacting proteins in mammalian cells." J Cell Biol 196.6 (2012): 801-810. PubMed PMID: 22412018...peptide (LPETG). In EXCELL, the mgSrtA variant can be displayed on the surface of a cell of interest, where...

... produced. Cells - Antibodies can also be produced in cell cultures using hybridomas, cell cultures created...created by fusing B cells with myeloma cells (immortalized B cells). The genetic reorganization that creates...identify and sort cells from a mixed population. Overview of application Collect cells from cell culture or ...specificity occurs in the B cell itself prior to antibody production; therefore, each B cell only produces one ... specific hybridoma cells into plasmids. Plasmids are then transfected into cells and induced to express...determines the isotype of the antibody. B cells, the immune cell responsible for producing antibodies, can...mature B cell can clone itself to enable higher production of a single antibody, but each B cell can only...

...engineered cells in therapy. The lungs are difficult, but other tissues like the retina have cell-based therapies...Chapter 9: Using stem cells as in-vitro models of disease. How Darrell is using stem cells from patients to...sharing reagents, cell lines, data, and knowledge without exclusivity. Chapter 11: The Stem Cell Bank (CReM ...the lungs, and, among other projects, using stem cells to develop in vitro models for lung disease. Darrell...what drives him as a praticing physician and stem cell researcher. Darrell's ultimate goal - to develop...means to treat disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...projects - differentiating surfactant producing cells that can be used to better understand the biology...

...like microbes in a drop of pond water or the cells in your cell culture dish. Microscopes come in a huge ...transmission electron microscopes that allow us to see cellular components with science-fiction levels of detail...

...transfected cells, we count the number of tag-positive cells (tag ICC) as a percentage of total cells in the...Deliver csRNP into cells: Prepare cells for transfection, add csRNP and ssODN to the cell suspension, transfect...csRNP plus ssODN, is then transfected into stem cells. Cells are allowed to recover for 2-5 days and then...efficiently these components are delivered into cells; for each cell type, the transfection/delivery step needs...Proteins - the workhorses of the cell – never work alone in the cellular milieu. It is, therefore, critical...DNA) to perform diverse biochemical tasks in the cell. One of the most popular approaches to study protein...with its associated protein complexes - from the cell-lysate using antibodies against the target protein...

...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...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 ... replaced by the gene of interest. As a result, cells that retain the ccdB gene die off and a higher percentage...

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

...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...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-...heavy chain, and they are small and stable inside cells. Nanobodies binding GFP were designed in 2009. Subsequent...

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