We narrowed to 591 results for: cel.2;
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TypeBlog Post...fusion location, there are many choices to make. 2. Prime Editing: Adding Precision and Flexibility to...introduction of nucleic acids into bacteria or eukaryotic cells is a common technique in molecular biology and scientific...
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Gamma-Retroviral Vector Guide
TypeGuide...range of infectivity for different cell types (known as tropism). Figure 2: Gamma-retroviral plasmids. Created...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or...-activated cell sorting (FACS) is used to sort cells expressing GFP and later, sorted cells are expanded...actively dividing cell types, whereas gamma-retroviruses can only infect mitotically active cell types. This...Using Packaging Cell Lines Gamma-retroviral vectors can be packaged using helper-free cell lines (Figure...-retroviral packaging cell line developed by Garry Nolan at Stanford. These cell lines contain gag-pol...and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for the infection of mammalian cells). Using... -
Antibodies 101: Fc Effector Functions
TypeBlog Post...affinity and specificity. Trends in Immunology, 29(2), 91–97. https://doi.org/10.1016/j.it.2007.11.004 ...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... -
Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation
TypeBlog Post...bioengineering 94.2 (2006): 322-329. PubMed PMID: 16528756. 2. Oh, Steve KW, et al. "Substantial overproduction ...at bioRxiv. AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral...of the packaging cells (Vandenberghe, 2010). Rather than being released into the cell culture supernatant... particles associate with the cell membrane and are taken into the cell via endocytosis. In our study,...AAV2 viral particles by our packaging cells by treating the cells with the endocytosis inhibitor sucrose...target cells is blocked following sucrose treatment. Moreover, several reports have shown that cells treated... yields approximately 1.8-fold without affecting cell viability. This effect appears to be serotype specific... -
Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight
TypeBlog Post...light sensing proteins. Furthermore, during the first 2 weeks of life, larvae already exhibit distinct behaviors... light sensitive proteins (opsins) to manipulate cell activity, enables researchers to silence or incite...channels that allow charged particles to cross the cell membrane when exposed to light. Neuronal firing ...channel LiGluR both cause ion movement that results in cell depolarization. They therefore function as a neuron...in negative chloride ions which hyperpolarize the cell. It therefore functions as a neuron “off switch....They found that photostimulating the Kolmer-Agduhr cells was sufficient to trigger the symmetrical tail movement...forward swimming. Furthermore, only Kolmer-Agduhr cells on one side of the body required photostimulation... -
Understanding GPCR Signaling Using TRUPATH
TypeBlog Post...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and... to environmental conditions, cells must be able to detect extracellular stimuli. One way they do so is...bound to the intracellular surface of the GPCR. Binding of a ligand to the extracellular surface of the...complex converts a signal received by a GPCR on the cell’s surface into action that can result in the initiation...number of signaling cascades, inducing different cellular responses. One of the biggest advantages of such...protein transducers. To use the TRUPATH system, cells are transfected with plasmids expressing the GPCR... and the appropriate Gγ-GFP2 and Gβ constructs. Cells are then treated with the luminescent enzyme substrate... -
Easing the Protein Purification Process with pCri
TypeBlog Post...PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary T. Britton, and Anne S...doing some research, notably determining the subcellular localization and the number of disulfide bonds... -
CRISPR 101: Drug discovery and CRISPR
TypeBlog Post...development and therapy. Nat Rev Drug Discov (2017) 16(2):89-100. DOI: 10.1038/nrd.2016.238 Additional resources...identification of potential targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens...knock out, etc. – CRISPR can be used to generate cell line or animal models which have the precise disease...Since CRISPR has been adapted for use in primary cells, many tissue types, and across species, it enables... phenotype. CRISPR can also be used to generate cell line knock-ins of patient mutations which are variants...CRISPR has revolutionized the generation of knockout cell lines which has made this control step extremely...in a cancerous background and treat the parental cell line and the X knockout with your drug(s). If a ... -
Some Like it Hot: Thermostable GeoCas9
TypeBlog Post...remained active in up to 30% plasma. Figure 2: Human plasma longevity of SpCas9 vs GeoCas9. SpCas9...mammalian cells. Harrington et al tested GeoCas9’s editing skills by transfecting HEK293 cells with GeoCas9...choice for a number of model organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs... 45 °C 50 - 70 °C Genome Editing in Mammalian Cells? Yes Yes Longevity in Serum?* No Yes Table ...experimental results. GeoCas9 RNP editing in mammalian cells Besides being able to take some heat, GeoCas9 can... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog Post... gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development...PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of regulatory T cells in ovarian carcinoma...activity, and consists of other cell types, especially immune cells and cells of epithelial origin. Suppressing...turns off T cell function upon interaction with APCs. Furthermore, tumor cells alter their cell-surface proteome...molecules needed for T cell effector function. Cancer cells also produce programmed cell death ligand (PD-L1... activity of CTLs and NK cells. These cells would normally kill cancer cells but are directed not to through...to use whole cell therapeutics consisting of various types of immune cells. Dendritic cells (DCs) are particularly... -
Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility
TypeBlog Post...Almo, Institute for Protein Innovation 1:18:45 - 2:01:18 (end) - Panel Alex Tucker (Ginkgo Bioworks)...their curation of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to ... characterised antibodies against all human extracellular proteins with community-based validation. They... -
Finding Your Science Policy Path
TypeBlog Post...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...example proposals and consider applying for their accelerator for some hands-on guidance. Check out my webinar... -
Special Delivery: Fluorophore Targeting for FRET Studies
TypeBlog Post...establishing the dynamic range of your FRET assay (Figure 2). These labeling methods are freely interchangeable...PMID: 20161833. PubMed Central PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding...Mammalian Cells pCMV-DnpK dinitrophenyl hapten Mammalian Cells pANAP AnapRS Mammalian Cells Note:... Qi, and Marcel P. Bruchez. "Advances in chemical labeling of proteins in living cells." Cell and tissue...endogenous biomolecules in cells nonspecifically, and they cannot cross cell membranes, so they are best... ion channels, or cell surface receptors) or with experiments on permeabilized cells. Biarsenical labeling...these compounds cross cell membranes, thus enabling FRET measurements in intact cells. The Gradia Lab has... -
Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands
TypeBlog Post... ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...fluorescent labels. In Nature Methods (Vol. 19, Issue 2, pp. 149–158). Nature Research. https://doi.org/10.1038... is technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to... Dyes that can pass through cell membranes can prove useful for live-cell imaging. Experimental context...Additionally, cell permeability is a critical factor, as it influences the dye's ability to enter cells without...enabled the visualization of both organic tissues and cell cultures, opening biological interiors to many inquisitive...fluorophores to enable highly specific visualization of cellular components, typically a protein of interest. Fluorescent... -
Safe Port in a Storm...How Addgene is Weathering the Pandemic
TypeBlog Post...majority of deposited materials to study SARS-CoV-2 would be fully open and available to industry requestors...they grasped the crucial role Addgene plays in accelerating research. Yet this was still not enough to ensure...type of biomedical science research. Our work accelerates and enhances productive collaborations as well...distribution of plasmids for COVID-19 research is accelerating essential research in laboratories all around...scientists worldwide with Addgene’s traditional excellence through whatever lies ahead this year. We can... -
Sensing Neuronal Dopamine
TypeBlog Post...developed fluorescent-protein based DA sensors (Figure 2). The dLight1 dopamine sensor In 2018, Lin Tian’s ...the brain has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011). This technique... across the semipermeable membrane from the extracellular fluid into the perfused liquid. The probe then...detection of dopamine in flies, fish, and mice." Cell 174.2 (2018): 481-496.PubMed PMID: 30007419. PubMed... -
Hot Plasmids and Viral Preps - July 2021
TypeBlog Post...method relies on the fluorescence ratio between the 2 fluorophores, rather than on the fluorescence intensity...Fluorescence Activated Cell Sorting (FACS), allowing for enrichment of successfully targeted cells that can be used...investigating genetic mechanisms related to morphogenesis, cell fate specification, and human disease. Sun,... -
RUBY-Red Siliques
TypeBlog Post...make it easier to study these mutants. Fig. 2: (A) Betacyanin is produced from tyrosine by the activity...graduate student studying plant development and cell-cell communication in the Nimchuk Lab in the Biology...untransformed seedlings or callus (undifferentiated plant cell culture), which is great for a wide variety of applications... -
Tag Your Favorite Yeast Genes with Ease
TypeBlog Post...Pringle’s former lab at UNC Chapel Hill. Bähler et al.(2) describe a modular collection of plasmids for a wide...insertion. The linear PCR product is introduced into the cell by direct transformation. A given insert will typically... -
Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes
TypeBlog Post...E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles...limit of detection for arg1 expressing ECN is 109 cells/ml, which is within the range of the ~1011 commensal...