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  1. Deciphering the Mysteries of Behavior with Viral Vectors

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    Blog Post
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    Sept. 17, 2019, 12:40 p.m.
    ...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...the mesocorticolimbic system in addiction. Front Cell Neurosci. 2014;8:466. PubMed PMID: 25653591. PubMed...
  2. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
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    July 30, 2024, 1:15 p.m.
    ...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...As part of Addgene’s 20th anniversary celebration, we’ve been finding and sharing stories unique to our... be safely used to deliver a genetic payload to cells, and the complexity of effectively deploying them... can be to use. AAVs, which can target different cell types depending on their serotype, are particularly...
  3. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
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    Nov. 3, 2016, 2:30 p.m.
    ...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools... the 926 video abstracts identified. The journal Cell had the largest average view count per video abstract.../wesharescience.com/pin/7472 Article: http://www.cell.com/current-biology/abstract/S0960-9822(16)30873...
  4. Antibodies 101: Beyond Surface Labeling

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    Blog Post
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    June 27, 2024, 1:15 p.m.
    ...which would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore...population.     Figure 2: Dump gating labels all unwanted cell lineages with the same fluorophore...out all CD8+ T cells from a mix of CD8+ T cells, CD4+ T cells, B cells, and dendritic cells. In this case...+ T cell, dendritic cell, CD8+ T cell. Created using biorender.com.   Intracellular labeling Oftentimes...in flow cytometry are cell proliferation and cell viability. Unlike intracellular labeling, dye staining...be done on live cells. For cellular proliferation, esterified dyes that can enter cells freely are typically...amines on the cell surface, which yields a much weaker signal on live cells compared to dead cells. As a result...
  5. Let There Be LITE Plasmids

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    Blog Post
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    Nov. 21, 2013, 3:56 p.m.
    ...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and... vectors and genetically target them to specific cell populations. They demonstrated use of the LITEs ...non-invasively, its use is not restricted to cultured cells but extends to freely moving animals, as established...novel mode of optogenetic control of endogenous cellular processes." The LITE system can now be applied...no means limited to particular model organisms or cell types. “This tool is really flexible and modular...
  6. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
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    Sept. 28, 2023, 1:15 p.m.
    ...chosen SspB-tag-GEF and your cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control ...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ... target to specific subcellular regions, laser ablation can damage other cellular structures, and genetic...demonstrated its use to probe cell mechanics, vesicle transport, and cell division, but given the long ... expressed in a monolayer of endothelial cells, showing cell protrusions and overlaps (arrows) after blue...2016). Rho GTPase signalling in cell migration. Current Opinion in Cell Biology, 36, 103–112. https://doi.org...Optogenetics gives you the power to control cells or organisms with the flip of a switch. You may be ...
  7. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

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    Blog Post
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    Sept. 9, 2025, 1:15 p.m.
    ...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...stereotaxically, STX) to the brain. Figure 2: Selection, prep, and testing of AAV enhancers in ...genetic access to cortical cell types. Cell. https://doi.org/10.1016/j.cell.2025.05.002 Hunker, A. C.,...Although all cell types share the same DNA sequence, not all enhancers are functional in every cell. Differential...expression in a cell type-specific manner also makes them useful tools for targeting those specific cell types ...Brain Map portal, and cell science-focused tools at the Allen Institute for Cell Science. There are also... primate neocortex. Cell Reports, 34(13), 108754. https://doi.org/10.1016/j.celrep.2021.108754 Panigrahi...
  8. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
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    Aug. 23, 2018, 12:05 p.m.
    ...microdomains of live cells." Science 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. ...phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2 coat...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...
  9. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
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    Sept. 14, 2016, 2:30 p.m.
    ...using a 2-step PCR-based protocol and are compatible with single-end or paired-end read flow cells. FREQ-Seq...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...Purification Process with pCri Cell-free protein expression kit Cell-free expression systems have been...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of...testing and comparison of different cell-free expression systems; the Cell-free expression test kit represents...: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq PUF Assembly...facilitate gene expression in bacteria, yeast, mammalian cells, insects, and even Xenopus oocytes. The expression...
  10. QC Sequencing Technologies at Addgene

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    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...short-read sequencing are highlighted in Figure 2. Figure 2: Overview of short-read sequencing technology...named) “nanopores” that are attached to the flow cell membrane and have an electrical current running ...
  11. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
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    Oct. 8, 2020, 1:15 p.m.
    ...caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma Contamination in Cell Cultures by Polymerase...bringing in environmental contaminants.  Figure 2: An Addgenie works in the tissue culture hood while...mycoplasma contamination in your cell culture or you are about to begin a new cell culture project. If mycoplasmas...present in most of the cell culture facilities, tissue culture labs and every cell culturist has to deal...cost-effective (Sung, 2006). The sensor cell-based method uses specialized cells that can detect the presence of... should be made available for cell culturing and maintenance of the cell lines. Providing such an environment...Photo by Kate Harten DeMaio. Cell lines and reagents Examine your cell line stock for mycoplasma contamination...
  12. Antibodies 101: Validation

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    Blog Post
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    March 24, 2022, 1:15 p.m.
    ...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...look at the subcellular localization of a protein of interest. Conveniently, you found a cell line in which...antibody binding in the transfected cells but not in control cells. But (there is always a ‘but’), you...transfect it into your favorite cell line. In parallel, you transfect cells with another plasmid not encoding...the cells transfected with your protein of interest and show just the faintest band from the cells transfected...when you start using complex samples such as whole cell lysate or tissue sections. It is important to confirm...
  13. Antibodies 101: The Basics of Western Blotting

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    Blog Post
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    March 8, 2021, 2:41 p.m.
    ... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of...readout for subcellular fractionations, a procedure that isolates proteins from different cellular compartments...interest from Addgene, transfected it into your target cells, now what? How can you tell if the protein you are... are so keen to study is expressing in your cells? Immunoblotting or simply the western blot, or western...specific protein within the complex mix derived from cells or tissues. Westerns are useful not only to detect...protein levels relative to standards, detect the cellular location of proteins, and can be a readout for...
  14. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
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    July 12, 2016, 2:30 p.m.
    ...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA... into a mouse embryonic stem cell (ESC) line, enrich and select for cells that had successfully incorporated... when it gets exciting – the cell sends out an “SOS” signal and cellular repair mechanisms rush in to ...Cas-Mediated Genome Engineering. Cell 154:1370–1379 . https://doi.org/10.1016/j.cell.2013.08.022 Yoon Y, Wang ...Cas-Mediated Genome Engineering. Cell 153:910–918 . https://doi.org/10.1016/j.cell.2013.04.025 Zhang X, Liang...develop into chimeric mice (with a fraction of the cells in the adult mice derived from the engineered ESCs...
  15. Six Spooky Science Stories and Halloween at Addgene

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    Blog Post
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    Oct. 31, 2019, 1:06 p.m.
    ...Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169...attaches to the cell wall of green algae (Soo et al., 2015). There, it divides into two cells. At the same...Vampirovibrio). What’s left of the algae cell is just the cell wall and cytoplasmic membrane. Vampirovibrio...
  16. Microbes: Look, Listen, and Tell

    Type
    Blog Post
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    Aug. 28, 2018, 12:04 p.m.
    ...similar optical quality (magnification of 140X and 2 micron resolution). I have worked many hours in a ... to microbes by using non-invasive methods for cellular imaging with ultrasound. In his research he applies...
  17. Top 10 Open Science Developments of 2013

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    Blog Post
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    Dec. 19, 2013, 3:46 p.m.
    ...access to the results of federally-funded research. 2)      In June, the White House honored 13 “Champions...Change,” including our recent guest Drew Endy, in celebration of open science. 3)      Also in June, the G8...
  18. 6 Steps to Submitting a Resume That Gets Seen

    Type
    Blog Post
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    July 9, 2015, 2:30 p.m.
    ...you are having trouble preparing your most excellent resume. 2) Follow job boards and check them regularly...give you a chance at an interview:  1) Have an excellent resume ready to be submitted The structure and...successful application. It takes time to prepare an excellent resume…it is worth it. Read here to get help if...
  19. What's New in CRISPR - March 2020

    Type
    Blog Post
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    March 10, 2020, 1:15 p.m.
    ...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout...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...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 ...
  20. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
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    Aug. 4, 2015, 2:30 p.m.
    ...photoconvertible fluorescent protein. Nat Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication... are you’ve used a GFP derivative to conduct subcellular localization studies or make a reporter construct...shown to work in plant, Drosophila and mammalian cells, and it may find wide use in biological research...turnover rate. In many cases, however, a GFP-tagged cell line or organism has already been made, and it would...made by accident. After exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et al. ...spontaneous conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White circles...this technique may not be appropriate for high-pH cellular compartments. Sattarzadeh et al. have shown that...
  21. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
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    May 12, 2015, 1:32 p.m.
    ...Chinese hamster ovary (CHO) cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin...recombination varies for different constructs and cell types. Second, Cre may induce recombination at pseudo...standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding domain-fusion...recombination efficiency. When expressed in neuronal cells, each line showed broad recombinase-dependent RFP...these recombinases were also active in mammalian cells. In experiments, B3 and KD were shown to be functional... damage induced by Cre recombinase in mammalian cells. Loonstra A, Voojis M, Beverloo HB, Al Allak B, ...recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity regulation. Heidmann...
  22. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

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    Blog Post
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    Jan. 21, 2020, 2:30 p.m.
    ...might otherwise be hard to identify. Figure 2: Users can cross-interrogate screens from the literature... disease, cell type or drug treatment can browse through screens performed in relevant cell lines or conditions... currently houses over 890 CRISPR screens in 629 cell lines, each manually curated and annotated from ...feasibility and power of CRISPR-based library screens in cell lines from humans and other species. By accurately... screens performed in 26 different breast cancer cell lines. Screenshot updated 2024. Each screen ...screen is captured by BioGRID curators, including the cell line/tissue type, enzyme system, sgRNA library and...example phenotype screens with read-outs other than cell viability. The following video provides an in-depth...
  23. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
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    July 11, 2017, 2:30 p.m.
    ...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to... the single cell level), are separated by, for example, cell sorting. The separated cells are subsequently...more than 8x107 successfully targeted cells. Depending on the cells used and the readout, this could be ...My group works on immune cells in different contexts. Most of the immune cells we study are migratory, ...CRISPR modified cells are transferred into an animal and the behavior of the transferred cells is followed...with an easily measureable cellular phenotype (e.g. survival of a cancer cell as a cytotoxic drug is added...
  24. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
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    Oct. 4, 2016, 2:30 p.m.
    ...closer to the clinic (Tsai and Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...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...
  25. New CRISPR Web Resources and #12DaysofCRISPR Recap

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    Blog Post
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    Dec. 12, 2017, 3:13 p.m.
    ...Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed...predict that we’d be able to edit RNA in mammalian cells with CRISPR? I certainly didn’t, but now we can ...updates We hope you enjoyed the #12DaysofCRISPR, a celebration of our updated blog resources. Not on social ...Improve HDR Efficiency for CRISPR Editing in Human Cells Day 3: Cpf1: Cpf1: A New Tool for CRISPR Genome ...
  26. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...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...
  27. Antibodies 101: Fc Effector Functions

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    Blog Post
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    May 18, 2023, 1:15 p.m.
    ...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...
  28. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    Blog Post
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    Dec. 13, 2018, 1:14 p.m.
    ...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...
  29. A Tour of Addgene's Most Popular Pooled Libraries

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    Blog Post
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    Nov. 19, 2025, 2:15 p.m.
    ...knockout cells can be used to identify genes that underlie a particular phenotype. Figure 2: Schematic...single-cell level. CRISPR libraries CRISPR screening has become a popular tool for probing cellular pathways...barcodes allow researchers to identify specific cells — as well as their descendants. This makes them ... for lineage tracing, among other applications. Cells can be identified using fluorescence, like in the...pooled library. Created with BioRender.com. The CellTag libraries from the Samantha Morris lab use a combinatorial...barcoding approach to observe clonal dynamics and cell lineage. Variations of this library allow tracking...genome-wide knockout screens in mouse and human cells, respectively. Last month, the David Root and John...
  30. Easing the Protein Purification Process with pCri

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    Blog Post
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    June 19, 2015, 3:08 p.m.
    ...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...
  31. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

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    Blog Post
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    July 16, 2019, 12:59 p.m.
    ...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...
  32. Understanding GPCR Signaling Using TRUPATH

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    Blog Post
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    Dec. 14, 2020, 2:15 p.m.
    ...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...
  33. CRISPR 101: Drug discovery and CRISPR

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    Blog Post
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    May 4, 2023, 1 p.m.
    ...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 ...
  34. Some Like it Hot: Thermostable GeoCas9

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    Blog Post
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    Sept. 14, 2017, 12:40 p.m.
    ...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...
  35. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
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    March 14, 2017, 2:30 p.m.
    ... 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...
  36. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

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    Blog Post
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    May 18, 2018, 7:32 p.m.
    ...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...
  37. Finding Your Science Policy Path

    Type
    Blog Post
    Published
    Sept. 22, 2020, 1:15 p.m.
    ...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...
  38. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...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...
  39. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
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    March 21, 2024, 1:15 p.m.
    ... ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...small-molecule fluorescent labels. Nature Methods, 19(2), 149–158. https://doi.org/10.1038/s41592-021-01338... 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...
  40. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
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    Sept. 17, 2020, 1:15 p.m.
    ...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...
  41. Sensing Neuronal Dopamine

    Type
    Blog Post
    Published
    July 30, 2019, 12:59 p.m.
    ...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...
  42. Hot Plasmids and Viral Preps - July 2021

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    Blog Post
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    July 27, 2021, 1:15 p.m.
    ...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,...
  43. RUBY-Red Siliques

    Type
    Blog Post
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    March 1, 2022, 2:15 p.m.
    ...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...
  44. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    Published
    Nov. 19, 2013, 2:37 p.m.
    ...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...
  45. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    Published
    June 19, 2018, 1:38 p.m.
    ...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...
  46. Viral Vectors 101: Transductions

    Type
    Blog Post
    Published
    April 30, 2024, 1:15 p.m.
    ... last ~2-7 days at moderate levels. With regard to target cells or organism, some primary cells and differentiated...packaging plasmids into a viral production cell line, usually HEK-293 cells. When all of these viral plasmids ...secreted into the cellular media, making viral collection fairly easy. Producer cell media can be collected... rogue HEK-293 cell). This virus-containing media can then be placed on whatever cells you would like ...just zapping your cells? That really depends on two things. First, what type of cell or organism context...process of transferring foreign nucleic acid into a cell by a viral agent. Transductions are similar to transfections...processes which introduce foreign nucleic acid into cells by any means, but are unique in that they require...
  47. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ... temporal activation of a transgene. Figure 2: FLEX Cre-Lox system.  WPRE The Woodchuck Hepatitis...about synthetic promoter AAVs for cell-type expression in retinal cells Resources on Addgene.org Find ...
  48. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    Published
    Aug. 2, 2018, 12:56 p.m.
    ...PMID: 30083608. PubMed Central PMCID: PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent...on freeze-dried, cell-free (FD-CF) components. Making proteins outside of a cell - cell free extracts Typically... into a host cell (through a process called transformation), essentially giving the cell all the instructions...to dispose of the live cells. The alternative is to express the protein in a cell-free manner. This is ...biology, especially those at the molecular and cellular level. The reason for this is cost and complexity...electrophoresis to separate DNA or transforming cells with green fluorescent protein, these activities...transcription and translation. Unfortunately, having live cells in a classroom presents equipment needs, such as...
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