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We narrowed to 1,028 results for: ELL

Showing: 561 - 600 of 1028 results

  1. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

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    Blog Post
    ...gives a talk at the Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and Beyond meeting...contains the deoxyadenosine deaminase, TadA-7.10 (Gaudelli et al., 2017). The idea was to evolve a TadA-7.10...III product is produced and can infect bacterial cells. If a phage does not deliver an active variant, ...passage from the infected to a fresh uninfected host cell culture). This dilution step represents the major...but gets manually passaged from an infected host-cell culture to an uninfected culture which results in...efficiently and in more contexts. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson...

  2. CRISPR 101: Non-Homologous End Joining

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    ...Translocations in Human Cells Are Generated by Canonical Nonhomologous End-Joining. Molecular Cell 55:829–842 . ...Genome Editing Specificity. Cell 154:1380–1389 . https://doi.org/10.1016/j.cell.2013.08.021 Waters CA, Strande... chromosomal DSBs. It is important to know that cells possess two major repair pathways – Non-Homologous... steps Unlike HDR, NHEJ is active throughout the cell cycle and has a higher capacity for repair, as there...should take less than an hour, thus a population of cells constitutively expressing a targeted Cas9 should...CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video] Pooled CRISPR Libraries Offer Genome-Wide...

  3. Visualizing Protein Turnover In Situ

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    ...proteins in cells. Homeostatic regulation of protein abundance is essential for normal cell function, and...one particular protein in neurons or in any other cell type. However, we cannot distinguish new from old...facilitate the visualization of specific proteins in cells. Tagged protein can easily be overexpressed from...techniques to follow translation in living single cells. However, before CRISPR-Cas9 these techniques were...interests, suspected to interact, are added to a cellular sample. Both antibodies have a short antibody ... turnover of specific endogenous proteins in any cell type. Exciting! Many thanks to our guest blogger... how small molecule fluorophores can be used for cell labeling Check out other protein tagging strategies...

  4. Giving gRNAs a Facelift - Synthetic and Beyond

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    ...gRNA technology have enhanced targeting efficacy as well as the unnecessary stress of worrying about the ...evolved ways to prevent degradation of important cellular RNA molecules with post translational modifications...deoxynucleotides. A partially DNA gRNA is surprisingly well-tolerated by the Cas9 protein and can increase on-target...slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing).  Some...transfection efficiency and even track localization in cellular experiments. There are many suitable dyes to choose...enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol 33, 985–989 (2015). https://doi....Enhance CRISPR Genome Editing Systems in Mammalian Cells. Frontiers in Genome Editing, 28 (2021). https:/...

  5. Antibodies 101: Conjugation

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    ...and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell Biol 89:e115. https://doi.org...need a little help to be useful. Mainly because, well, antibodies are kind of hard to see. To solve this...imaging instruments or X-ray film and lends itself well to assays like western blots and ELISAs. Chromogenic... like proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et...throughout antibody sequences, up to 80 per antibody (Mueller, et al., 1988). If some of those lysines are in...doi.org/10.1002/cpcb.115  Mueller BM, WrasidloO WA, Reisfeld RA (1988) Determination of the Number of e-Amino...epitope and transcriptome measurement in single cells. Nat Methods 14:865–868. https://doi.org/10.1038...

  6. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    Blog Post
    ...sensitive GFP2 (roGFP2)-based biosensors for live cells. His work focuses on the enzyme hydropersulfide-... understand the processes that happen inside the cell,” says Pedre Pérez. Figure 2: Micrographs...Micrographs of the biosensor expressed in the human cell line, Hep3B. From left to right: Mitotracker Red, GFP...studies opioid use disorder. Her goal is to identify cell populations in the brain that contribute to the ...will explore the connectivity patterns of these cells and the functional contribution to a variety of ...administration and withdrawal to explore how these cells may encode the negative affect associated with drug...library of mutant sensors expressed in a mammalian cell line engineered to be electrically excitable. The...

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

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    ...is non-pathogenic, well-studied and inexpensive. In addition, a complete set of well-annotated deletion...contain methionine. (This allows cells carrying nonfunctional as well as functional MET genes to grow.... or students clone similar sequences from a less-well studied organism into overexpression plasmids. Students...synthesis in S. cerevisiae (Figure 1) occurs through a well-characterized pathway with multiple enzymatic steps... 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...

  8. Design Tips for Prime Editing

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    ...editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652.e29. https...Guide RNA and Target DNA. Cell, 156(5), 935–949. https://doi.org/10.1016/j.cell.2014.02.001 Yan, J., Oyler-Castrillo...type of edit, genomic context, delivery strategy, cell type, and goal of the experiment. Now, onto the ...nicking of the unedited strand and encourage the cell to use the edited strand as a template during DNA...also allows certain unintended edits to escape cellular surveillance. To limit unintended incorporation...https://doi.org/10.1016/j.cell.2021.09.018 Nelson, J. W., Randolph, P. B., Shen, S. P., Everette, K. A...

  9. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    ...depends on its spike protein to bind ACE2, the host cellular receptor. To more easily study the spike variants...protein, called HexoPro, was developed by Jason McLellan’s lab. This version of the spike protein was developed... investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids express these...scientists used the genome-wide CRISPR libraries, Brunello (knock out) and Calabrese (activation), from Addgene...for SARS-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their search, they found...packaging) and contain an IRES-Puro marker for stable cell clone selection. For more information on these plasmids...Lavinder JJ, Ippolito GC, Maynard JA, Finkelstein IJ, McLellan JS (2020) Structure-based design of prefusion-...

  10. 6 Tips for Grant Writing

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    Blog Post
    ...to give yourself plenty of time when writing a fellowship proposal. Yes, yes, I hear you saying “We know...research (either because of expertise, equipment, or excellence of the institution). Likewise, if you’re staying.... Become a science communicator More and more fellowship applications are asking for a 200-300 word explanation...explanation of your research for non-scientists. Most fellowships are funded by the general public (either through...may make your perfectly reasonable paragraph unintelligible to a lay audience. My lay paragraph typically...assisting me with applications for much larger fellowships. Also, if you were awarded money to carry out...Cambridge University where he studied mechanisms of cell division. Recently Seán got the entrepreneurial ...

  11. The Materials Science of Optogenetics Experiments

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    ...currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the Rockefeller University.  His interests...protein-coupled vertebrate rhodopsin in cerebellar Purkinje cells. The Journal of biological chemistry. ...Pharmacology and Optogenetics. Cell 162.3 (2015): 662-674. doi: 10.1016/j.cell.2015.06.058. Pubmed PMID: 26189679...Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University of Michigan...PMCID: PMC3466483. 4. Kim TI, et al. Injectable, cellular-scale optoelectronics with applications for wireless...

  12. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ... genetic loci within a single cell. One might imagine transfecting cells with both the nuclease-positive...Perhaps most importantly, most Cas9 orthologs are less well characterized than the conventionally used SpCas9... both to lower relative nuclease efficiencies as well as to more stringent targeting rules that result...nuclease-competent Cas9, obviating the need to generate new cell lines or transgenic animals expressing dCas9 as ...reliance on orthogonal Cas9 species. Within a single cell, full-length guides targeting one set of genetic...nanoparticles as drug delivery systems. She is also a NASA Fellow studying biosignature preservation in Mars analogue...knockin mice for genome editing and cancer modeling." Cell 159.2 (2014): 440-455. PubMed PMID: 25263330. PubMed...

  13. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

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    ...produced by a single B cell. To create a monoclonal antibody cell line, myeloma cells are fused with activated...activated B cells from the spleen of mice injected with the antigen of interest. The immortalized cells, called...distributing over 3,000 antibodies and associated hybridoma cell lines. The DSHB was created by the National Institute... individually isolated and expanded. The clonal cells are screened to identify hybridoma lines, each producing...

  14. A Scientist's Guide to Ebola

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    Blog Post
    ... infected cells to go undetected by the immune system in a novel way, by shielding host cell surface proteins... itself to escape vesicles within host cells and turn the cell into a veritable virus factory.  Their ... sets off.  The virus works its way into immune cells in ways that aren’t fully understood. Much of the... viral attachment and fusion. Once inside a host cell, new evidence in PLoS Pathogens shows that the Ebola...have information or another resource to add, please tell us about it in the comments. ...

  15. Inntags: Innovative Protein Epitope Tagging

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    ...indicated that inntags IT5, IT6, and IT10 expressed well in cells and did not form undesirable aggregates. Using...tags. Impressively, IT6-GFP did not reduce yeast cell size or viability compared to untagged GFP overexpression...multiple other proteins. Again, little to no change in cell morphology was observed with IT5 and IT6 fusions...and HA tags had especially detrimental effects on cell size and growth rate, indicating that they compromised... replace our older epitope tags - only time will tell! References 1. Georgieva, Maya V., et al. "Inntags...correlation for protein localization in mammalian cells." Nature methods 10.4 (2013): 315-323. Pubmed PMID...

  16. Plasmids 101: Codon usage bias

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    Blog Post
    ...bodies that will generally segregate with the cell pellet during purification procedures. Even if you produce...codons at random and expect your sequence to express well in any organism. So what are the evolutionary constraints...reasons include: Metabolic pressures - it takes cellular resources to produce tRNAs that recognize different...Tuller et al. found that translation efficiency is well correlated with codon bias in both E. coli and S...’t necessarily mean the gene is going to express well. If you do get good expression, you should also ...mismatched translation rates and potential effects on cell growth, even hosts producing alternative tRNAs may...a means to fine-tune gene expression." Molecular cell 59.2 (2015): 149-161. PubMed PMID: 26186290. PubMed...

  17. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    ...Syndrome and Related Disorders. Cell 152:984–996 . https://doi.org/10.1016/j.cell.2013.01.038 Guy J, Gan J, ...deliver the precise amount of protein to each brain cell to restore physiological levels. Much research is...repair a gene in a living animal across multiple cell types using a mouse model for a human neurological... that enough wild-type MECP2 is made inside the cells, but this is exciting progress in tackling the root...Resource Center. Researchers can also find links to cell line repositories including patient-derived iPSCs...Modifications in Human-Induced Pluripotent Stem Cells. Front Genet 10: . https://doi.org/10.3389/fgene...Neurological Disorder by Programmable RNA Editing. Cell Reports 32:107878 . https://doi.org/10.1016/j.celrep...

  18. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    ...Generating a no-fuss genetic knockout  Knock-out cells or animals are engineered by introducing Cas9-mediated... which cuts at NRN PAMs with high efficiency, as well as NYN, but with a slightly lower efficiency. Information...transcription by blocking its initiation. In mammalian cells, robust repression requires targeting dCas9 along...study essential genes whose knockout would cause cell lethality. The Krüppel associated box (KRAB) is ...repressing target genes in a variety of species and cell types can be found on Addgene’s website. Activating...activating target genes in a variety of species and cell types can be found on Addgene’s website. ...optimized for expression in different species and cell types including (but not limited to) bacteria, yeast...

  19. Important Considerations in Optogenetics Behavioral Experiments

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    ...currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the Rockefeller University.  His interests...optogenetics, ICSS can now be done with neuronal cell-type and temporal precision. In ICSS, a test chamber...refine and optimize your stimulation parameters as well. Inhibition of neuronal activation is a little ... remote contextual memories. This study is an excellent example in which the authors started with a brain... research. Derek P. Simon received his PhD in Cellular and Molecular Biology from the University of Michigan...differential participation in reward circuitry." Cell reports 8.6 (2014): 1857-1869. Pubmed PMID: 25242321...Dynamics of retrieval strategies for remote memories. Cell. 2011;147(3):678-89. Pubmed PMID: 22179551. Pubmed...

  20. Plasmids 101: E. coli Strains for Protein Expression

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    ... want to make sure your cells have first reached an appropriate density. Cells in the exponential growth...collecting dead cells, and, conversely, you cannot induce too early as there are not enough cells in the culture... purposes, these E. coli strains are not usually well suited for recombinant protein expression. Many ...host protein, using up valuable resources in the cell thus leading to slowed growth. Additionally, some... promoter. Although the DE3/T7 RNAP system works well for most experiments, the lac promoter can “leak...such as TB or 2XYT is optimal because of the high cell-density they support; however, minimal media supplemented...

  21. Fluorescent Proteins 101: History of Fluorescent Proteins

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    Blog Post
    ...crystal jellyfish Aequorea victoria (Figure 1) in molecular detail. On the underside of the “umbrella” of ... chemiluminescent proteins, like luciferases, as well as fluorescent ones, like Green Fluorescent Protein...green glimmer, Shimomura collected many, many jellyfish specimens from Puget Sound in the Pacific Ocean...he was able to isolate two proteins from the jellyfish’s photoorgans; the first, which he called aequorin... could be used to measure gene transcription, as well as to track protein localization. And as it turns..., another part of the fluorescent protein puzzle fell into place, as they, taken alongside the insights...

  22. Plasmids 101: Expressing shRNAs for RNA Interference

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    ...transfection or viral delivery of a plasmid. RNAi is an umbrella term for a number of different applications that...miRNA and piRNA can be found endogenously in many cell types, while shRNA and siRNA are synthetic constructs...methods. shRNA processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the...shRNAs is to generate stable knockout or knockdown cell lines. In fact, it was the go-to choice for many...shRNAs are a quick way to create stable knockdown cells lines, and you can usually find validated shRNA ...Resources References Bartel, D. P. (2004). MicroRNAs. Cell, 116(2), 281–297. https://doi.org/10.1016/s0092-...Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998). Potent and specific genetic interference...

  23. CRISPR 101: Drug discovery and CRISPR

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    ...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 ...research field! References and Resources References Fellmann, C., et al., Cornerstones of CRISPR-Cas in drug...the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the...

  24. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

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    ... 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...

  25. Addgene at the ASM Microbe 2016 Conference

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    ...plasmid sharing services, as well as #happydepositors who stopped by to say hello. Needless to say we all ...booth. As well as being the Director of the Great Lakes Bioenergy Research Center and a fellow Rhodobacter... plenty of geeky fun to be had at ASM Microbe as well as the serious business of science. Two symposia... work this Broad Institute lab is doing creating well-optimized CRISPR screening technology. Addgene got...making popular screening libraries like Brie and Brunello available to the scientific community. However...

  26. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

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    ... a look! mNeonGreen: a yellow-green fluorescent protein mNeonGreen is a yellow-green fluorescent protein...protein that was derived from the tetrameric yellow fluorescent protein (LanYFP) from the marine invertebrate...was reported as the brightest monomeric green or yellow fluorescent protein at the time. It is 1.5 to 3...mNeonGreen wavelengths lie between the green and yellow fluorescent protein wavelengths, it can be imaged...with standard green fluorescent protein filters or yellow fluorescent protein filters, with no or minimal...a FRET donor to mCitrine and mOrange or for live cell imaging of fusions and FRET reporters. mTFP1 is ...

  27. Antibodies 101: Epitope Availability

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    ... have been extracted from a cell, instead of when it is attached to a cell membrane.  Picking the Right...even in that mythical place where antibodies are well-organized, documented, and easy to find (though ...partners, the epitopes available for binding change as well. If this change hides the epitope that the antibodies...with native proteins instead of denatured ones - well, don’t relax just yet. Proteins often change shapes...epitopes that are available on the outside of the cell. The same protein may have different epitopes available...need to validate on denatured proteins or on live cells. But for others, you may need to validate that it...

  28. CRISPR 101: Off-Target Effects

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    ...millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted cells to sequence...trying to generate a knockout cell line, with the goal of isolating single cell clones to use for multiple...generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation arising.... a level of, say 5%, then the majority of single cell editing events will be on target. Thus, if you isolate...example, if your goal is to generate a knock-out cell line to perform downstream characterization of, ...experiments on is a good way to control for off targets as well as clonal differences. If you are performing CRISPR...

  29. A Control for All Seasons

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    ...to detect dead or dying cells so that you can base your analysis on living cells only (for example, propidium...Fluorescence Microscopy.” Journal of Cell Biology, vol. 185, no. 7, Rockefeller University Press, June 2009, ... positive control group, with an alternative but well-studied kinase known to prevent tau hyperphosphorylation...interactions will be observable in preimmune serum ‌as well as with an isotype-matched control.      ...This tends to be most problematic when staining cells or tissues that are the same species as your primary...propidium iodide only stains dead cells). Compensation controls address the spectral overlap that occurs...

  30. "Build Your Plasmid": The Game - Play to Teach

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    ... of human stem cells when students are not even allowed to manipulate prokaryotic cells? Especially, if...should be “brains-on”, but as much as I try being compelling, interactive, and application-driven to boost...where you need to apply basic molecular biology, cell biology, microbiology, genetic engineering and strategic...could be used to genetically manipulate human stem cells. Then they had 10 minutes to expose their strategy...construct and explain why they selected a certain target cell population. Who is the winner? Ultimately, society...gene therapy in the following lessons. And let me tell you more, they had brilliant ideas on how to solve...

  31. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

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    ...Awardees: Xin Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their... is currently a graduate student in Thomas J. Mansell’s lab at Iowa State University where she is developing...reliably detect HMOs. Fatima’s first design is a whole cell biosensor with two components - an enzyme that breaks...proposes using this sensor to detect HMOs found on cellular surfaces. After incubating the sensor with bacteria...binding results in an increase in fluorescence of cells that actively produce the HMOs. Fatima has demonstrated...work at her home town in Bangladesh where it was well received. In addition to her research endeavors,...out Fatima's recent publication on her HMO work in Cell Chemical Biology! Additional Resources on the Addgene...

  32. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

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    ... glutamate release from neurons and other brain cells in vivo. Glutamate plays a variety of roles in synaptic...(SF-Azurite-iGluSnFR), green (SF-iGluSnFR), and yellow (SF-Venus-iGluSnFR). iGluSnFR3: Improved Glutamate...photostability, quantum yield, and expression. Create yellow (v82) and lime green (v857) variants differing ... versatility through two different variants The yellow (v82) and lime green (v857) versions can be summarized...sensor to the membrane and displays it on the extracellular side. In addition to the original PDGFR TMD,...promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin-1 promoter. Good for neuronal...promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter, FLP-dependent ...

  33. Scientists Map the SARS-CoV-2-Human Interaction Network

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    ... used HEK293T cells in our first study, but we're planning to soon expand to other cell types, including...26 of the 29 SARS-CoV-2 viral proteins in human cells and used these viral “bait” proteins to fish for...human proteins that the virus interacts with in cells. Because viruses rely on the help of human proteins...monkeywrench into the virus’ hijacking of human cells. Even better, more than half the drugs hitting these...they have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a fairly... as the EU, Russia, India and Saudi Arabia -- as well as closer to home in the US and Canada. In fact,...

  34. 5 Great Apps for Lab Life

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    ...’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new...spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find an app that could...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...tools are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution...educational section. This section holds the Gibco Cell culture manual and basic instructional videos. The... Overall, it’s a great tool for someone starting cell culture work. 3. NEB Tools by New England BioLabs...alternative, similar apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme...

  35. Synthetic Photobiology: Optogenetics for E. coli

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    ...Marcotte EM, Voigt CA, Ellington AD. Cell. 2009 Jun 26;137(7):1272-81. doi: 10.1016/j.cell.2009.04.048. PubMed...with sophisticated gene expression controls. Most cell types do not respond to low-intensity light, so ...bacterial camera that prints a chemical image, as well as a genetic method for the computational problem... are compatible with many strains of E. coli, as well as with other plasmid-based systems, which will ...ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte EM, Voigt CA. Nature. 2005 Nov 24...

  36. Advice for Scientists Starting a Lab

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    ...carefully select first hires who are well-suited, productive, and fit well with your goals. These first hires...and foster an effective working environment? Tom Ellis was offered a start-up package that included a postdoc... most surprising thing about becoming a PI? Tom Ellis has a one word answer: “Email. Nothing prepared ...differently or the same as your adviser did? Tom Ellis merges the influence from his two PhD advisers who...our latest results as soon as the gels ran.” Dr. Ellis tries to lead his team as a scientists and a friend... lab!”   Acknowledgments Thank you to Dr. Tom Ellis (Imperial College London), Dr. Connie Cepko (Harvard...

  37. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

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    ...editor in mammalian cells Not all Cas13d proteins are functional in mammalian cells, but Konermann et al... Ruminococcus flavefaciens (nicknamed CasRx), as well as EsCas13d. Find the Yan et al. plasmids here ...make it possible to target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding...natural diversity of CRISPR systems, only time will tell how large the genome and transcriptome engineering... with RNA-Targeting Type VI-D CRISPR Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: ... a WYL-Domain-Containing Accessory Protein.” Mol Cell. (2018) pii: S1097-2765(18)30173-4. PubMed PMID:...

  38. A Deep Dive into BioBuilder

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    ...want to learn more about Synthetic Biology. Can you tell me where to start?” is a question more and more ...2011, BioBuilder is the brainchild of Dr. Natalie Kuldell from MIT’s Department of Biological Engineering...from her 20 years of college teaching experience, Kuldell sought to create a program that would capitalize...contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom are...plan or outstanding test scores.  As a result, Kuldell launched the BioBuilder Educational Foundation ...of Professor Richard Murray. Murray’s lab uses cellular extracts to perform transcription and translation...develop and test hypotheses. Students then use cellular extracts as a means to rapidly prototype DNA circuits...

  39. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    ...Drosophila cell line, I’ve studied a transcription factor important for T cell differentiation (a cell type ...integration will have eyes that are anywhere from light yellow to red. The variation in color depends on how transcriptionally...vermillion (another gene controlling eye color) and yellow (a gene affecting the body color of the adult fly...regeneration and homeostasis in the Drosophila midgut. Cell. 137, 1343–55 (2009). PubMed PMID: 19563763. PubMed...et al. Digoxin and its derivatives suppress TH17 cell differentiation by antagonizing RORγt activity. ...

  40. Multiplex Genome Editing with CRISPR-Cpf1

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    ...expression and needing to work with a transfectable cell line. 2) They require larger expression vectors... of 4 crRNAs in vitro and when expressed in 293 cells. This allows for a single promoter to drive expression... were used.  With transfection, 6.4% of HEK293T cells had edits at 4 of 4 targets when an array of 4 crRNAs...transfected on a single plasmid. This is versus 2.4% of cells transfected with a pool of plasmids that contained...processing of the crRNA array when expressed in cells, but notice how Zetsche et al reverses the orientation...mouse neurons and in vivo. For these experiments, cells were infected with a 1:1 ratio of two AAVs,  one...RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3), 759-771. PubMed PMID: 26422227. PubMed Central...
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