We narrowed to 762 results for: cell

...difficult-to-transfect cell types ranging from primary to embryonic stem cells. Viral integration of a... also generally less toxic to cells than transfections, preserving cell viability. Library delivery Screening...Viral applications Viruses work by infecting a host cell (the target of viral infection) and delivering a...to be anything you want delivered to your target cells. This delivered DNA/RNA can then be either permanently...permanently integrated into the genome of the target cell or transiently expressed. It’s important to note...This vector can express your construct in target cells transiently or constitutively depending on the virus...transcription factors, barcoded libraries to trace cell lineage, and so much more! Libraries allow you to...

...4,000 cell lines. Seventeen of these cell lines were used to investigate the susceptibility of cells from...additional cell line used in this study came from the Japanese Collection of Research Bioresources Cell Bank...that SARS-CoV, which entered human cells via hACE2, could not enter cells expressing murine ACE2 (Subbarao...repositories that distribute mouse strains, plasmids, cell lines, and much more have been critical for pandemic...from Reproducibility for Everyone. Case study 1: Cell lines Many of the reagents used to investigate SARS-CoV...accelerating research into this virus. One such example is cell lines. The American Type Culture Collection (ATCC...of this pathogen. The sharing and distribution of cell lines from repositories on opposite sides of the...

...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... the bathing solution and require uptake by the cells. Which of these strategies works best for a specific...showing that its acid tolerance is maintained in cells.   GFP is big GFP is a 28 kDa protein that resembles... This approach enables the study of proteins in cells, but not other biomolecules of interest e.g. DNA...several components that are naturally present in cells or media. These sources of ‘autofluorescence’ include...

...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...

... where many different cell types, or even different phenotypes of the same cell type, are intermingled...postsynaptic cells when used at very high titers (Zingg et al., 2017), which can lead to cells in far away...experimental conditions and measurements; (2) the cells you’re targeting; (3) how much expression is needed... capsids, which determine how a virus enters the cell. Different serotypes utilize a number of different...co-receptors, and other mechanisms to gain entry into the cell (Dudek et al., 2018; Korneyenkov & Zamyatnin, 2021...2021), meaning what works great in one system or cell type can completely fail in a different system (...one serotype that will efficiently transduce your cells. In fact, we usually recommend testing more than...

...reagents and keep the cells happy. Harvesting virus Once you have transfected your cells, wait 2–5 days and...however, you’ve collected the producer cells, you’ll need to lyse the cells to let the virions out. We recommend...particles that can transduce cells, which can vary based on target cells and can be altered by freeze/...come into contact with the rAAV and apply common cell culture practices diligently throughout the production...capsid genes.  In this helper-free system, HEK293T cells, expressing Ad E1A and E1B, can be transfected with...and then collect the supernatant and/or producer cells for further processing. What component(s) you need...released into the culture media, while others remain cell-associated (Vandenberghe et al., 2010). For certain...

...packaging AAV is transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids...precipitation. Instead of sonicating the cell pellet, I lyse the cells with this AAV lysis buffer and freeze...bath to release the virus particles from the cells. Cell lysates are then treated with benzonase immediately...five days after transfection, the AAV-containing cells and media are harvested and purified. In total, ...not counting the time needed to expand the HEK293 cells. Check out the Addgene AAV Production protocol for... strains, like NEB Stable. NEB Stable competent cells lack RecA, a protein that aids in the homologous... AAV. I skip the PEG precipitation of the HEK293 cell culture supernatant because it’s time consuming ...

...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...timers, researchers can look at a single image of a cell and understand how protein localization changes ...was a tetramer with a propensity to aggregate in cells (Tsuboi et al.), limiting its use. Nonetheless, ...versatile timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus embryos. Of course...understand the relationship between the age of a cell, protein, or cellular structure and a particular...location, start of gene expression, development of a cell structure, etc). FP timers should therefore find...

... for stem cells, or metastasized tumor cells, or alzheimers-affected neurons, or [your cell line of interest...critical to stem cell survival. Knowing which genes influence the survival of our cell culture is the foundation...minimal side-effects on healthy cells and maximal effect on unhealthy cells. Until recently, the human genome...ENCODE project, find enhancers in embryonic stem cells, and conclude with a targeted CRISPR screen that...essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds just over 13,000 significant...We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus...The result is a heterogenous population of stem cells, usually housed in a single flask. Through a bit...

...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... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...monitor gene expression from that promoter in a given cell type. This type of transcription reporting was among...find optogeneic actuators and sensors at Addgene. Cell marking/selection: Expression constructs like plasmids...

... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ...plasmid to generate a population of mutant cells. The mutant cells were treated with WNV at a dose and duration...regulate the ability of WNV to induce cell death in infected cells (SEL1L, UBE2J1, EMC3, EMC2, DERL2, UBE2G2...AID-tagging system to essential genes in human cells and mouse ES cells because it was difficult to fuse the AID...-to-study genes in hard-to-study cell types. Natsume T, et al. Cell Rep. 2016. PubMed PMID: 27052166 ...variety of phenotypes, including but not limited to cell growth and drug resistance. Recently, the Wu Lab... genes involved in West Nile Virus (WNV) induced cell death. The Wu lab library consists of 77,406 individual...

.... Nat Cell Biol. 2022. https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal...with one visual barcode per cell and up to 200-plex with two barcodes per cell. The researchers validated...long sequence knock-ins Visual barcodes for live cell clonal multiplexing Improving flexibility for ... powerful approach for proteomic mapping in both cell culture and whole organisms (see our 2019 overview...reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally have higher risks ...multiplexing by: Rachel Leeson Multiplexed images in live cells remains a challenge due to multiple factors, including...flourescent proteins that can be inserted into a cell without disrupting its functions. Recently, Ravid...

... 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...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...is very amenable to genetic manipulation. Yeast cells are great model organism not only for the reasons...

...Worms are particularly well suited for studies in cell biology, development, and aging. Scientists can .... elegans hermaphrodite consists of 959 somatic cells, which makes them attractive for imaging and fluorescence...researchers to express desired constructs in precise cell types and/or developmental timeframes. Here we highlight...

... of a microtubule-associated protein to regulate cell growth dynamics Article contributed by Klaus Wanisch...of the modified EB1 protein are overexpressed in cells, microtubule growth can be switched off with blue...paper demonstrated that the migration of a cancer cell line equipped with photodissociable EB1 could be...any genetic or pharmacological manipulations on cells, and has potential use for a wide range of applications... in cellular biology. Van Haren J et al. Nature Cell Biology 2018 Pubmed PMID 29379139. The “Riboglow...used to visualize RNA localization in mammalian cells. These tools consist of cobalamin-binding riboswitches...proteins (FP) with improved tissue penetrance and live cell imaging capabilities. Upon developing a new imaging-based...

...and inside the cells and are thus typically used for distinguishing live from dead cells. The combination...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...Another way to label proteins is through treating cells with fluorophore-conjugated antibodies that attach...

...tissue tropism, allowing them to target particular cell types. Figure 1: Diagram of AAV components...key that lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic...change how the viral particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular...multiple coreceptors, which may be present on different cell types and sometimes vary between target species ...strains. Pro tip! Getting a vector into a particular cell type depends largely on the viral capsid, but specificity...transduce skeletal muscle, cardiac, and retinal cells (Issa et al., 2023). AAV2 Despite being isolated...muscle, lung, CNS, kidney, retinal, and pancreatic cells (Ahuja et al., 2025; Issa et al., 2023; Wu, Asokan...

...mammalian cells. Here, we will cover how these structures are generated, why it’s important for cells to sense...performs almost all of its biological functions in our cells in the single strand form, but double stranded RNA...with bidirectional transcription. In an uninfected cell, dsRNA rarely circulates, but upon viral infection...Unlike dsRNA, DNA:RNA hybrids are abundant in normal cells and are arise from a ssRNA engaging one strand of...genes is constantly occurring throughout a single cell, so these nucleic acid hybrids are always around...at risk. Consequences of duplexed RNA Mammalian cells have multiple receptors in place to sense dsRNA ... dsRNA levels reach a given threshold within the cell, as is often the case upon viral infection. Endogenously...

... on the method used to introduce the gRNAs into cells. Most CRISPR pooled libraries have been created ...the chance of transducing a only a single gRNA per cell. The viral DNA containing the gRNA then integrates...next-generation sequencing of the genomic DNA. However, some cell types are poorly transduced by lentivirus, and thus...CRISPR libraries was not possible. Primary murine T cells cannot be transduced by lentivirus and have been...library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express Cas9 and screen...CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction, ..., now enabling forward genetic screens in cells with few genetic tools. Find the mouse CRISPR knockout...

...cancers. It was inspiring to participate in the 2017 Cell Symposium on Cancer, Inflammation and Immunity and...recent findings in basic immunology, cancer-immune cell interactions, cancer immunotherapy, as well as new...example the upregulation of the immunosuppressive cell surface receptor PD-1. This immunosuppressive environment...follow the interactions between tumor and immune cells in real time. Addgene depositor Professor Max Krummel...effect of PD-1 blockade.“ (Ref 4). PD-1 helps tumor cells evade the immune system - COX inhibitors like aspirin...therapeutics such as  (Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in ...combination of the CRISPR/Cas9 methodology with T cell receptor engineering (Ref 5)  as described by Blue...