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We narrowed to 587 results for: cel.2

Showing: 221 - 240 of 587 results

  1. Addgene Depositors Get More Citations

    Type
    Blog Post
    ... 1 2010 3 Nature  3.3% 10.5%   3.2  9.6 2 2010 167  Nature 3.3%  10.5%  3.2  534.1 3...Microbiology  1.6% 1.2%  0.75   125.3 In figure 2, you can see the full impact from using these weighted...plasmids deposited at Addgene still accrue on average 2 to 3 times as many citations as other papers in the... most relevant to plasmids: Genetics & Heredity, Cell Biology, Biotechnology & Applied Microbiology, Biochemistry...

  2. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...phosphorylation." Nature cell biology4.5 (2002): E127-E130. PubMed PMID: 11988757. 2. Hornbeck, Peter V., ...phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the importance of... the open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...optimized Sep-OTSλ and can recommend its use (6).  2. C321, a specialized bacterial host strain for pSer-incorporation...commonly cited that approximately one-third of cellular proteins are modified through phosphorylation ...translation of many stop codons and detrimental effects on cell growth. The C321.ΔA strain was genomically recoded...is particularly interested in the intersection of cell signaling, metabolism, and the mighty mitochondrion...

  3. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ... living plant cells." (2013). PubMed PMID: 24194740. PubMed Central PMCID: PMC3810607. 2. Bajar, Bryce... fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly to...PubMed PMID: 10500161. PubMed Central PMCID: PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators...fluorescent proteins available. Fortunately, several excellent reviews have collated data from various papers... alters the FRET efficiency. Detecting certain cellular changes with an intermolecular probe is often ...Miyawaki et al., 1997), was designed to measure intracellular calcium and published in 1997. Since that time...of labs to answer sophisticated questions about cellular processes. Currently, FRET experiments can probe...

  4. Chemogenetics Guide

    Type
    Guide
    ...muscarinic receptor and has not been used in vivo . Figure 2: DREADDs, their ligands, and signaling properties.... Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling...hM1Dq hM5Dq Human M3 muscarinic G αq CNO* Increase Ca 2+ Neuronal burst firing Armbruster et al., 2007 rM3D...channels, their effect, and outcome in neurons. Table 2. Descriptions of PSAMs and their activity in neurons...Forbes SL, Liggett SB (2001). Modification of the beta 2-adrenergic receptor to engineer a receptor-effector...targeted to specific tissues, cell types or even subcellular regions of a neuron. Cell-specific expression of...Human Stem Cell-Derived Neurons Repair Circuits and Restore Neural Function. Cell Stem Cell. 28:112-126...

  5. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ... the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating Gene function by... characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...specific cell types or entire organs in planarians. For example, the gene Piwi labels neoblast cells and the...techniques to alter gene expression or to label specific cells, but there are too few resources to teach students... in tissue. This is useful for labeling specific cell types and for viewing how gene expression changes...

  6. Lentiviral Vector Guide

    Type
    Guide
    ...intact. First-generation plasmids include (Figure 2): Transfer plasmid — contains transgene and wild-type...envelope removed Envelope plasmid — contains env Figure 2: First-generation lentiviral plasmids. Created with...plasmid 1 — contains gag and pol Packaging plasmid 2 — contains rev Envelope plasmid — contains env Third-generation...The inside out of lentiviral vectors. Viruses , 3 (2), 132–159. https://doi.org/10.3390/v3020132 PMID: ...Infectivity and Integration Targeting. PLoS ONE , 2 (12), e1340. https://doi.org/10.1371/journal.pone.0001340...features of lentiviral vectors. Human Gene Therapy , 24 (2), 132–142. https://doi.org/10.1089/hum.2012.229 PMID...production cell line, typically human embryonic kidney 293T (HEK293T) cells. Standard HEK293T cells are sufficient...

  7. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...lately (1, 2), depends on knowledge of the structure(s) a given phage will recognize on a host cell. To make...per plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now...microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and ... biology 34 (2015): 69-77. PubMed PMID: 26275959. 2. Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis... cloned into E. coli cells, isolated colonies will be lifted onto nitrocellulose membranes and exposed... proteins directly onto the nitrocellulose for subsequent probing for cell-binding activity.  First, cut...with live host cells Now you can proceed with incubating this membrane with your cells of interest and...

  8. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...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...

  9. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ... DC. 2. President's Council of Advisors on Science and Technology [PCAST] (2012). Engage to Excel: Producing...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...lack uracil, but contain methionine. (This allows cells carrying nonfunctional as well as functional MET... is available at our Investigations in Molecular Cell Biology Website. Many thanks to our guest blogger...right) has taught courses in biochemistry, molecular cell biology and genetics at Boston College. She is particularly...

  10. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down options for you...our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first... question about what should go into a cell, hover over the cell and instructions will pop up. When you... into the appropriate full and partial sequence cells. *Note: If you believe you need to deposit a collection...

  11. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific expression of molecular...specific cell type. Find some transduction characteristics of select AAV serotypes in Table 2 from the...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...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...

  12. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...digestion of the destination vector with SapI yields 2 sites- the first site accepts the sgRNA target sequence...marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the same type will produce... gRNA vector that targets the gene of interest, 2) a plasmid to specify the reading frame of the insertion...and function of cellular systems. FPs have been used to track protein localization, cell structure, intracellular...associated with cellular organelles, scientists have been able to study many cellular processes, including...to fluorescently tag markers of cellular structures in mammalian cells. These plasmids use fluorescent...Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors.” Cell Rep.15(1) (2016...

  13. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    ...pH visible at the sample interface upon thaw (Fig. 2). Before thawing, all the samples were at the control...showed that AAV infectivity was not altered by a short 2-hour exposure to pH as low as 2.5 (Potter et al., ...Methods. 2013 Apr;10(4):278-9. PubMed PMID: 23538862. 2. Potter M, Lins B, Mietzsch M, Heilbronn R, Van Vliet...measure the pH of our samples, we used a solution of cell culture medium, which is buffered and contains the...

  14. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ... of components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or ...overhang specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual... (KO, CRISPRa, CRISPRi, editing)   Figure 2: Selecting module fragments on the Fragmid website...Drosophilia Plasmid pRDB_051 (for stable fly cell lines; attL and attR sequences for genome integration...Drosophilia Plasmid pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration...rapid assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/...

  15. CRISPR Guide

    Type
    Guide
    ...Haploid Embryonic Stem Cells Carrying a Guide RNA Library. Cell Stem Cell . 17 (2), 221–232. PMID: 26165924...used to transduce your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either...of CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa... regulation of transcription in eukaryotes. Cell . 154 (2), 442–451. PMID: 23849981 Gilbert, L. A., Horlbeck...Transcriptional Programs with CRISPR RNA Scaffolds. Cell . 160 (1–2), 339–350. PMID: 25533786 Cas9 alternatives...single RNA-Guided endonuclease of a Class 2 CRISPR-CAS system. Cell , 163 (3), 759–771. PMID: 26422227 Zetsche...left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only...

  16. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ... Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038...single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227.   ... Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide specificities...particularly helpful for targeting non-dividing cells, which are difficult to modify through homology-... Cpf1 orthologs known to be active in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing... (RNPs) failed to induce off-target editing in a cell culture model. What we still don't know about Cpf1...reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016 Jun 6. PubMed PMID: 27272384...

  17. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...base pair to another base pair at a specific site. 2. Temporal, spatial, or conditional control of CRISPR...difficult-to-engineer Sulfolobus islandicus rod-shaped virus 2 (Mayo-Muñoz et al., 2018). Only viral particles that...Editing of Sulfolobus islandicus Rod-Shaped Virus 2. Viruses 10:695 . https://doi.org/10.3390/v10120695...Inhibitor of CRISPR-Cas9. Cell 170:1224-1233.e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha...Off-Switches for CRISPR-Cas9. Cell 167:1829-1838.e9 . https://doi.org/10.1016/j.cell.2016.11.017 Shin J, Jiang...concern is that the longer Cas remains active in a cell, the greater chances there are for off-target edits...within the first six hours. When added to human cells six hours after the introduction of Cas9 RNPs, AcrIIA4...

  18. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...resolved loci with physical map distances of 75 and 2 Mbp, with the calculated fluorescent distances correlating...established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that they could evaluate the...be applicable to studies of cell cycle progression, epigenetics and cellular reactions to external stimuli...Weissman JS, Qi LS, Huang B. Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001. Pubmed. Find...LS, Weissman JS, Vale RD. Cell. 2014 Oct 23;159(3):635-46. doi: 10.1016/j.cell.2014.09.039. Epub 2014 Oct...labeled Cas9 to label multiple genomic loci in live cells. While other systems can be used to label loci, ...CRISPR/Cas9’s ease of use and ability to label live cells make this system truly advantageous. This new technique...

  19. Tetracycline Inducible Expression

    Type
    Collection
    ...expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA-2 Contains Gateway L1L3 sites and EF1α promoter driving...systems were tested for functionality in mammalian cells. The lac system with the inducer isopropylthio-β-galactoside... and Bujard tested the tet system in a mammalian cell system (HeLa) and found that the tet system was ... interest rtTA or TetR expression plasmid Stable cell lines can be made that continuously express a system...derivatives can be viewed in Krueger et al, 2004. Some cell culture sera may contain tetracycline or its derivatives...of gene expression at a single copy in mammalian cells; Expresses firefly luciferase hairpin and GFP under...vector lentiviral Gateway RNAi system for conditional cell line generation; contains expression cassette for...

  20. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...Figure 2, following compensation. Figure 3: Flow plot for a double positive sample. The cell population.... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations.   Figure 2: Compensation example based on experimental sample... make it difficult to properly gate your desired cell population. By compensating between fluorescent ... your fluorescent markers are staining for. If a cell is positive for both markers, it will show up as...learning! Ashlyn has a research background in CD8+ T cells and cancer immunotherapies. She is currently a research...
Showing: 221 - 240 of 587 results