We narrowed to 714 results for: cel
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TypeBlog PostPublishedJan. 29, 2019, 2:27 p.m....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 ...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...as stable metabolite sensors for imaging in live cells. By placing Spinach downstream of a riboswitch, ...
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Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....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)....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... -
What Good Citizenship Can Do for Reproducibility in Science
TypeBlog PostPublishedNov. 7, 2019, 2:18 p.m....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... -
An Introduction to Adenovirus
TypeBlog PostPublishedOct. 3, 2023, 1:15 p.m.... 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...hexon), as well as other intracellularly-exposed proteins and downstream cellular events, all recruit strong...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....production and/or by genes present in the HEK293T cell line used (which have fragments of the AdV genome... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m....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...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...Hamster Ovary (CHO) cells. (L'Abbé et al., 2018). Recombinant antibodies can be produced through a single... -
Adenoviral Vector Production and Troubleshooting
TypeBlog PostPublishedJune 14, 2018, 11:44 a.m....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... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog PostPublishedSept. 21, 2021, 1:15 p.m....improved cell-free system to examine biosynthetic pathways A new tool for enriching ABE-targeted cells The ...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...multiplexed and can be used in human pluripotent stem cells, a cell type that has been difficult to modify in the... Corner New ready-to-use viral preps Improved cell-free transcription/translation system to examine...biosynthetic pathways By Gabrielle Clouse Recently, cell-free transcription/translation systems (also referred....0c00581 New tool for enrichment of ABE-targeted cells By Ellie Adams Single base modifications through... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog PostPublishedOct. 17, 2019, 12:28 p.m.... specific cell-type promoter. Once injected into the brain, the viruses infect all the cells with an inactive...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... This could be overcome by fusing opsin genes to cell-type specific promoters; however, those usually ...drive weak expression of downstream genes. Strong cell-specific promoters, on the other side, are too long... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog PostPublishedJuly 2, 2020, 12:40 p.m....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 ... -
Plasmids 101: Knockout/Knock-In Plasmids
TypeBlog PostPublishedDec. 1, 2016, 3:30 p.m....recombined cells. Recombination is a rare event, so you must select as opposed to screen for cells where recombination...Correctly recombined cells will not contain the negative selection marker, but cells with random recombination...modified version of the targeted sequence to the cell which will recombine the template with the endogenous... used to inactivate specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout...targeting construct. For recombination to occur in a cell, a minimum of 2 kb of sequence homology is required...antibiotic resistance gene. To avoid selecting for cells in which the construct has randomly integrated into...outside one of the homology arms. When we select cells later, we will first perform a positive selection... -
Lentiviral Vector Uses and Overview
TypeBlog PostPublishedMay 19, 2016, 2:30 p.m.... 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...ability to transduce both dividing and non-dividing cells. Lentiviral vectors are the vector of choice for...lentivirus HIV-1, well-known to infect non-dividing cells. The first lentiviral vector system consisted of...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 ... -
Viral Vectors 101: Types of viruses
TypeBlog PostPublishedJune 6, 2023, 1:15 p.m....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...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...lentivirus can infect both dividing and non-dividing cells. Lenti particles enter the nucleus through nuclear... -
Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME
TypeBlog PostPublishedJune 6, 2019, 12:41 p.m....efficient gene deletion in B cells, T cells, macrophages, and dendritic cells, with no deleterious effects...to HEK293FT and HeLa cell lines. When the miRNA activity profile for a given cell type is already known...or ZFN to edit immune cells poses challenges such as the inhibition of immune cell differentiation. Furthermore...immune cell activity. However, a CRISPR-based tool for in vivo functional analysis of immune cells has been...construct in bone marrow stem cells from Cas9-expressing mice, and used the cells to reconstitute the immune...post-transcriptional gene expression across the majority of cellular pathways. Despite several years of research, the..., these reporters can help define the different cells present in a mixed population, or identify differentiation... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog PostPublishedJan. 16, 2025, 3:42 p.m....particle sneaks its way into cells. Generally, AAV serotypes have a primary cellular receptor and sometimes ...transduction efficiency, and is an excellent choice for targeting cardiac cells (Bish et al., 2008). It can ... Therapy: An Overview. Cells, 12(5), 785. https://doi.org/10.3390/cells12050785 Kaludov, N., Brown, K....delivery across species. Cell, 184(19), 4919-4938.e22. https://doi.org/10.1016/j.cell.2021.08.028 Tervo, D...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...multiple coreceptors, which may be present on different cell types and sometimes vary between target species ... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog PostPublishedFeb. 8, 2018, 12:17 p.m....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...Promega supports the Addgene mission statement: Accelerate research and discovery by improving access to... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog PostPublishedOct. 19, 2017, 1:54 p.m....specific cell types (populations) within neuronal networks in living animals. To accomplish such cell specific...individual cells. With this technique, a Neuroscientist can see the full dendritic arbor of a single cell with...Cre-mediated possibilities for site specific (and often cell type specific) control of DNA recombination and ...30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular evolution...revolutionize neuroscience Manipulation of specific cell types in neuronal networks Double inverted vectors...combination with mouse lines that express Cre only in the cell type of interest (Atasoy et al. 2008; Cardin et ...single neuron and its synapses in the living brain? Cells of a specific brain area can be transfected by in... -
Supporting Rare Disease Research with the 2020 BeHEARD Awards
TypeBlog PostPublishedSept. 8, 2020, 2 p.m.... 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...Genomics Institute BeHEARD (Helping Empower and Accelerate Research Discoveries) Challenge provides grants...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... -
Sequencing Options for CRISPR Genotyping
TypeBlog PostPublishedOct. 4, 2016, 2:30 p.m....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...picture of the impact of CRISPR editing in the model cell line or organism. Several options exist for unbiased...Osborn et al. 2016) and HTGTS (Frock et al. 2015) for cell-based experiments. These strategies can be used ...off-target editing events. Two of the most common cell-based methods are genome-wide, unbiased identification... -
CRISPR in the Clinic
TypeBlog PostPublishedNov. 2, 2023, 1:15 p.m....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...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, ...therapies involve editing healthy donor-derived cells and delivering them to the patient. Current... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m.... 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...