We narrowed to 859 results for: ELL

.... FPs are generally bright, stable and well tolerated by cells. They can be used for protein localization...of neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in the...ability to interact with protein partners within the cell. However, antibodies can have low affinity for these...trigger protein aggregation which is toxic for the cells and affects the labeling of small structures like...team, an ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with...is a symbol of opposition to the teaching of intelligent design in public schools and was started in 2005...used to amplify the direct smFP signal within sub-cellular structures. For instance, smFPs have been used...

...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without... to date for visualizing genomic loci in living cells. Previous iterations of genomic loci visualization...labeling system to visualize chromosomal loci in human cells (Ma et al., 2015). To do so, they fused a different...different hairpins will produce a secondary color (yellow, cyan, or magenta) depending on the hairpin combination...visualization of sites on different chromosomes in living cells, the system requires high-copy chromosome-specific... aptamer system to visualize the gRNA in living cells. Broccoli aptamers are nucleic acid sequences that...production needed to deliver CRISPR-Sirius into cells. They named this guide sgRNA-Sirius-8XMS2 which ...

...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...

...mCherry for FACS sorting of the weaponized T-cells (CAR-T cells) - and then delivering them to a tumor - ...effectively as standard CAR-T cells, but it reduces the off-target effects caused by T cell migration.  CAR expression...Adapted from Fig. 2 Campbell et al, Nature Methods   Find YFP plasmids here!   Campbell, B.C., et al. Chemically...reducing the on-target off-tumor activity of CAR–T cells was recently devised by the Yingxiao Wang lab.  ...above can be made for expressing your favorite T-cell receptor, or perhaps any other tool that can benefit...  Wu Y, et. al. Control of the activity of CAR-T cells within tumours via focused ultrasound. Nat Biomed... is that some antibodies may just not perform as well in one application as another, so it is always best...

...-binding scFv onto a well-folded scFv scaffold to improve its stability in cells, and use it in a variety... including live-cell imaging, single-molecule tracking, immunolabeling of fixed cells, Western blotting...possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular Cell. 2021. https://doi.org...Anti-FLAG Frankenbodies for modular imaging of cellular targets Optimized genetic screening technique ...Anti-FLAG Frankenbodies for modular imaging of cellular targets by: Mike Lacy Adding to their set of anti-HA...frankenbodies to amplify the signal.  Liu et al. Genes Cells. 2021. https://doi.org/10.1111/gtc.12893, Zhao et... Cas12a for multiplexed gene perturbation by: Gabrielle Clouse While many CRISPR-based genetic screening...

...neatly between yellow and red fluorophores, allowing it to be paired with green, yellow, and/or red sensors... nerve and muscle function, hormone release, and cell signaling throughout the body. Because calcium levels...used simultaneously to monitor or control different cell types or signals in the brain. For this to work,...emission (solid) spectra for various teal, green, yellow, orange, and red fluorophores. The spectra for ...indicators jRGECO1a and jRCaMP1a are widely used as well. To complement these tools, we recently developed... researchers can image multiple brain regions or cell types at once using existing indicators. For example... using  iGluSnFR4 to monitor glutamate in green/yellow and RdLight to monitor dopamine in red.  However...

...can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants...transport studies in HEK cells, recombinant protein purification in bacterial cells, and a two-hybrid protein...for synthetic biology applications in bacteria or cell-free systems. The approach is particularly useful...tags to your proteins, then the CIDAR Kit is an excellent option (Iverson et al., 2016). CIDAR offers a ...add-on kit and is compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo ...plant-specific parts you‘ll need, including sub-cellular localization signals, plant markers, and reporters...of expression and integration needed in mammalian cell types, and comes with a large library of common ...

...training, so of course I felt compelled to participate in one of the satellite Marches for Science (my local...currently a post-doctoral fellow at Vanderbilt University. She studies beta-cell function and failure in... duty to tell our science stories in a manner that encourages open discourse with our fellow community...also incredibly impressed with how well organized the march was, as well as the friendly atmosphere that ...constant vigilance and actions beyond marches, as well as allies outside the scientific community. Vanessa...responsibility to join the public conversation and to tell the stories behind our research.  That's why I marched...against legislators and because the march had been so well-publicized, we might encounter hostile anti-science...

...intensity evolves over time in living cells at a power used for live-cell imaging. To conclude, the most photostable...molecule will photobleach at some point. For live-cell imaging, it is desirable to have fluorescent proteins...experiment. The best fluorescent proteins for live cell imaging can be excited many times, thereby producing...these factors, environmental conditions such as cellular redox state and oxygen concentration may affect...protein is a key requirement for quantitative live-cell imaging, it is important to be able to quantify ...it is advisable to perform time-lapse imaging of cells producing the fluorescent protein of interest under... a substrate. These in vitro methods allow for a well-controlled environment, but they do not mimic the...

...Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar S, Davies BW, Ellington AD, Moran NA, Barrick...Control of Gene Expression. Cell 152:1173–1183 . https://doi.org/10.1016/j.cell.2013.02.022 Ronda C, Pedersen... is primarily a means of selection that targets cells in which homologous recombination has not occurred...ensures high editing efficiency; the only non-edited cells to survive have inactivating mutations in the Cas9... E. coli, the system was used successfully in Tatumella citrea, another species of Enterobacteriaceae,...one or more genes in E. coli. and in a mammalian cell line (Qi et al., 2013). Two important features of...other Cas9 orthologs to identify a more efficient, well tolerated, Cas enzyme. They identified the Cas9 ...

...measuring protein expression or localization in cells, or even single-particle tracking of the bound complex...) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent protein tags. ...(1–10) and GFP(11) Versatile protein tagging in cells with split fluorescent protein. Kamiyama et al. ... and mNG3A(1–10) for use with mNG2(11) Improved yellow-green split fluorescent proteins for protein labeling...amplification. Zhou et al. PLoS One. 2020 Bo Huang Yellow Venus pBiFC-VN173, pBiFC-VC155 Identification...pairs with sfCherry2(11) Multiplexed labeling of cellular proteins with split fluorescent protein tags. ...

...Genome-wide Inhibition Library. DNA and cell amounts After deciding on a cell type, you will need to determine...DNA and how many cells to use for your transformation(s). The amount of competent cells you need total ... depending on the particular type of cells you are using, as well as other factors, such as your lab’s...generalized amplification protocols tend not to work as well.   Defining your library First, you’ll need to collect... generally electroporated into electrocompetent cells to ensure the highest possible transformation efficiency...efficiency. Using Stbl4 or other ultra-high efficiency cells ensures that plasmid diversity in the library is...transformations, such as splitting 400 ng of DNA in 100 μL of cells over four individual electroporations. The specifics...

...same: Deliver CRISPR library to cells of interest. Select for cells of interest (positive or negative...infect specific cell or tissue types. Using serotypes that have a known tropism for your cells of interest... a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing cells for long...mice are involved. Ex vivo starts with transducing cells with the CRISPR library in vitro before transplanting...interest — for example, AAV8 for kidney cells — can help improve infection rates in your experiment. ...AAV vectors, enhancing their affinity for specific cell and tissue types. The combination of serotypes and...integration sites seem to only occur in the target cells or tissues. AAV also shows improved transduction...

...degraded by the cellular machinery, leaving a small amount of active NgAgo/ssDNA complex in the cell. As proposed...results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL NgAgo ARTICLE... guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA ...dangerous, non-physiological temperature for mammalian cells and one which diminishes the endonuclease activity...guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid.... Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or...lipofectamine-based method to deliver NgAgo ingredients into the cell, the same method used by Gao et al, 2016 (1). View...

...large amounts of carbohydrates that can inhibit the cell lysis process. To ensure that lysis is complete,...strains adapted for protein purification use the cell’s resources to produce high levels of protein, not...difference? In the simplest of terms, more volume = more cells = more plasmid. Plasmids can vary in copy number...of plasmid copies supported within each bacterial cell in your culture. Copy number is partially specified...richer media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield... supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific...glucose (in moderation) may also serve to increase cell density and plasmid yield. Start with a single, ...

...them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially if you...reporters allow you to easily screen or select for cells that are expressing your gene of interest at high...promoter, multicistronic plasmids ensure that any cells that are positive for your marker should also be...anytime you want to express multiple genes in the same cell. Although it is possible to drive co-expression ...promotes translation of the viral genome in eukaryotic cells.1,2 Since then, a variety of IRES sequences have...discovered - many from viruses, but also some from cellular mRNAs. What they all have in common is the ability... cistron due to factors such as the experimental cell type and the specific genes cloned into the vector...

...Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific expression of molecular tools...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...with AAV: 1. Getting your viral vector into your cell of interest: AAV tropism and route of administration...transduction, the viral capsid proteins bind to the target cell’s surface proteins and this interaction determines...efficiency of the AAV serotype used for a specific cell type. Find some transduction characteristics of ...by restricting expression to genetically defined cell types. Specific transgene expression can be also...different approaches can increase the specificity of cell targeting even further. For example, injecting AAVs...

...for studying the bacterial cell cycle, asymmetric cell division, and cellular differentiation, but it takes...crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-based DNA demethylation Optogenetic repressors...complex remains in the cytoplasm. Read the paper in Cellular and Molecular Bioengineering Find the optogenetic...Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image...popular Streptococcus pyogenes system does not work well in Caulobacter). To test their CRISPRi system, the...lab repressed ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and...piggyBac transposon system for human pluripotent stem cells Lindy Barrett’s lab developed new fluorescently ...

...already available today to cure sickle cell anemia in stem cells but this can’t be used in people until...and how much we have yet to learn. As Dr. Mello said, “Cells have been in the information age for billions...meeting movie stars, but when I met the astronaut Jim Lovell I was speechless (the club of people who have been...genome engineering, Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier. I was joined by Addgene Scientist...from them during a panel discussion with Dr. Craig Mello and Dr. Paul Stoffels from J&J. The panel was moderated...the mechanisms of RNAi, the field for which Craig Mello and Andrew Fire won the Nobel Prize. What did Dr... audience discussed how J&J is already modifying cell and animal genomes to test hypotheses of disease...

...used. The Fujii lab demonstrated this in mammalian cells by expressing dCas9 with an N-terminal 3xFLAG tag...studies (Fujita et al., 2018). In addition to the "in cell" form of this method, in which an engineered DNA-binding...molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii lab also developed another...Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification with an anti-Cas9 antibody... disease for which there is currently no cure (Campbell et al., 2018). Find plasmids from the Fujii lab...see our interview with Dr. Fujii. References Campbell, Amy E., et al. "NuRD and CAF-1-mediated silencing...recombinant CRISPR ribonucleoproteins." Genes to Cells 21.4 (2016): 370-377. PubMed PMID: 26848818. Fujita...