Skip to main content

We narrowed to 813 results for: cell

Showing: 401 - 450 of 813 results
  1. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ..., or express the insert they carry in the target cell. These vectors allow you to test the function of...If you are planning to track your protein in the cell or purify it – and you do not have a specific antibody...selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent fusion ...get away with generating transiently transfected cells, you will not need a mammalian selection cassette.... Other vectors can be expressed in fly culture cells like S2 and carry the metallothionein promoter (...
  2. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...temperature incompatible to genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus...can block Cas12a-mediated genome editing in human cells. Read the paper in Science Find the anti-CRISPR...anti-CRISPR expression plasmids CRISPR-based method for cell lineage tracing Valcyrie is a CRISPR-based marker...marker excision method for marking distinct cell clones developed by the Michalis Averof lab. It’s an alternative...
  3. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ... primary cells. The authors used this platform to assay dozens of GPCRs across many cell types in...biosensor platform. Cell, 187(6), 1527–1546.e25. doi: https://doi.org/10.1016/j.cell.2024.01.028. New...Prime editing (PE), initially developed in human cells, has been uniquely challenging to apply in dicotyledonous...labeling approach using these constructs in HEK 293 cells (Smith et al., 2024). They then used single-molecule..., CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only...recordings of spontaneous activity in human stem cell-derived neurons expressing ASAP5-Kv. C) Cultured...
  4. MXS Chaining

    Type
    Blog Post
    ...construct was then introduced to HeLa cells (Figure 1). The resulting HeLa cells show robust labeling, with strong...protein localization, as well as cellular processes like gene expression and cell migration (Sladitschek and... MXS-chaining were used to label subcellular structures in live cells. Four individual expression constructs...imaging or flow cytometry applications in mammalian cells. The modules used in this method include fluorescent...components, one can generate constructs to visualize cell cycle dynamics, titrate inducible transgene expression...applications in flow cytometry approaches in mammalian cell culture systems (their original goal). Their work...detection of each fluorophore at the respective subcellular structure (Figure 1). Here, MXS Chaining gave...
  5. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...(FRET) microscopy imaging of live cell protein localizations. J Cell Biol. 2003 Mar 3;160(5):629-33. ...measurement of single molecule interactions in living cells using a microscope. Historically, molecular interactions...suitable for studying interactions within living cells. By coupling fluorophores to proteins, pioneers ...directly detect protein/protein interactions in living cells. Since then FRET has also been used to measure conformational...efficiency and cross-talk. Methods to Measure FRET for Cell Biology Studies Several methods have been used over...microscopy: monitoring protein interactions in living cells. Bioessays. 2012 May; 34(5):341-50. doi: 10.1002... Constructs to target mTurquoise2 to various subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian...
  6. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...producer cell line, such as HEK-293T, production of viral particles, and transduction of a target cell line...re derived from viruses that can infect people, cells, and animals. Some of these viruses are completely...While the virus can still enter (and thus infect) cells, this safety feature prevents the virus from replicating...replicating and spreading in non-target cells, including yourself as the researcher.   Figure 1:... the procedure. Mice move much more quickly than cell cultures do, and tired researchers are accident-...vector carries an shRNA to a gene essential for cellular function, this would also be a higher risk vector...
  7. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    ...proteins that bind an RNA motif of interest in living cells. Challenges with existing RNA-protein interaction...and naturally occurring complexes present in the cell, it can also introduce non-physiological RNA-protein...a short timescale, such as during drug action or cell signaling. Limitations of RaPID Like other BirA*... controls include both scrambled RNA motifs and cells transfected with BirA* protein alone (i.e. no RNA...protein that’s highly expressed in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels...2018). RNA–protein interaction detection in living cells. Nature Methods, 15, 207-212. PubMed PMID: 29400715...
  8. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...chemically. Enzymatic biotinylation occurs naturally in cells and is the process by which biotin is covalently... Immunosorbent Assay) Southern/Northern blotting Cell surface labelling Attaching polypeptides to a substrate...substrate Flow cytometry/fluorescence-activated cell sorting (FACS) In situ hybridization Methods for...proteins in bacteria, yeast, insect and mammalian cells. Check out Kay et al., 2009 for methods on biotinylating...Application of avidin-biotin technology to improve cell adhesion on nanofibrous matrices.” Journal of Nanobiotechnology...purification of tagged transcription factors in mammalian cells and transgenic mice.” PNAS 100.13 (2003):7480-5....-H., Lee, C.-Y., & Li, P.-C. “Buoyancy-Activated Cell Sorting Using Targeted Biotinylated Albumin Microbubbles...
  9. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ... of human stem cells when students are not even allowed to manipulate prokaryotic cells? Especially, if...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...
  10. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ... of all of the mRNA bound-ribosomes in a cell. When cellular mRNAs are digested with ribonucleases, ribosomes...the effects of 5’ UTR variants in translation of cell cycle protein Emi1, showing that ~80% of transcripts...powerful tools for studying translation in various cell types and physiological states. References 1. Ingolia...imaging the first round of translation from single cells to living animals.” Science 347(6228) (2015): 1367...of Translation of Single mRNA Molecules In Vivo.” Cell 165(4) (2016): 976-89. PMID: 27153498 PMCID: PMC4889334...Regulating translation is key to cellular function, especially during development or stress. With ribosome...explore the complexity of translational control in cellular physiology. Ribosome profiling Ribosome profiling...
  11. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ... between Cas9 (which causes the breaks) and host cell DNA repair factors (which repair those breaks) makes...example, correcting the allele that causes sickle cell anemia. However, HDR occurs much less frequently...a structure could drive sequence replacement in cells. Targeting three dCas9 molecules to a precisely ...functional Cas9 protein and the targeting guide RNA into cells by electroporation. Results from our lab and others...Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins...
  12. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ... yeast, plant, insect, and mammalian cells, including human cells. piggyBac is biased to integrate at ...is active in vertebrates and integrates in human cells at rates similar to retroviral vectors. The hyperactive...piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive PB...transposase (hyPB) is ~10-fold more active in mammalian cells compared to the codon-optimized wild-type piggyBac...G/C)TNA. Tol2 can deliver 10-11 kB to mammalian cells without decreased efficiency, with a maximum cargo...and has lower efficiency of integration in human cells than piggyBac and Sleeping Beauty. Minimal Tol2 ... or barcodes. When transposons are delivered to cells or a model organism, they insert into the host’s...
  13. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...can be used in bacterial and cell-free expression systems, mammalian cells, yeast, flies, and even plants...transport studies in HEK cells, recombinant protein purification in bacterial cells, and a two-hybrid protein...for synthetic biology applications in bacteria or cell-free systems. The approach is particularly useful...add-on kit and is compatible with E. coli and/or cell-free systems. Protein expression in yeast MoClo ...of expression and integration needed in mammalian cell types, and comes with a large library of common ...protein interaction study in yeast cells. Need more MoClo? Check out Addgene's MoClo Guide! Customizing...Construction of Biological Circuits in Mammalian Cells.” ACS Synthetic Biology, vol. 8, no. 11, American...
  14. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...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...
  15. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ... be introduced into host cells, such as bacterial, yeast, or mammalian cells, for antibody production ...Nanobodies and recombinant binders in cell biology. Journal of Cell Biology 209:633–644 . https://doi.org...animals and bleed dates and even clonal hybridoma cell lines can produce more than one mAb (Bradbury et...plasmid or from an integrated sequence in a stable cell line. The genes encode the heavy chain and light...antibody protein binds an antigen. Image from Fvasconcellos.     How are Recombinant Antibodies Made...
  16. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable for up to...possible and submitted our manuscript to the journal Cell just six weeks later. The Zika publication timeline...of Toronto. His research is focused on developing cell-free synthetic biology applications for human health...Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from...de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939. PubMed PMID: 25417166. PubMed...Keith, et al. "Paper-based synthetic gene networks." Cell 159.4 (2014): 940-954. PubMed PMID: 25417167. PubMed...
  17. CRISPR Plasmids - Epigenetics

    Type
    Collection
    ... time and are potentially heritable in dividing cells. CRISPR activation and interference technologies...
  18. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...Xenopus frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then ...is the smallest photosynthetic cell in existence, but most abundant cell on earth (twice the human biomass...Hawaiian bobtail squid has an extreme number of nerve cells and an unusually high level of RNA recoding in the...using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all (maybe because...total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little bits of hydra can...Mormyridae (Jason Gallant) Weakly electric fish are excellent systems to study for convergent evolution because...
  19. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...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...
  20. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...Generating a no-fuss genetic knockout  Knock-out cells or animals are engineered by introducing Cas9-mediated...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... have delivered Cas9 and a sgRNA to your target cells, it is time to confirm that your target sequence...
  21. Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants

    Type
    Blog Post
    ...expressing target cells. Any neutralized pseudoviruses would be unable to get into the target cells and this can...disease severity and survival. Cell 184:476-488.e11. https://doi.org/10.1016/j.cell.2020.12.015 Garcia-Beltran...vaccine-induced humoral immunity. Cell 184:2372-2383.e9. https://doi.org/10.1016/j.cell.2021.03.013 Additional resources...vaccines and allows the virus to gain entry into human cells via human ACE2 receptors. In their assay, the pseudoviruses...
  22. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...have become indispensable tools for molecular and cell biologists over the last two decades, but their ...quantum yield, and when transiently expressed in cells, generates much less signal than comparable fluorescent...three luminescent proteins can be used together in cells. As Nano-lanterns are not subject to photobleaching...Nano-lanterns, Takai et al. continuously imaged cells for about 4 hours! The latest enhanced Nano-lanterns... al. “Luminescent proteins for high-speed single-cell and whole-body imaging.” Nat Commun. 3 (2012): 1262...light. Long-term light exposure can also lead to cellular phototoxicity, and experimental success can be...phototoxicity, they successfully imaged multiple subcellular compartments over several minutes using Nano-...
  23. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced cells. Grant...in the Mammalian Genome. Cell 167:233–247.e17 . https://doi.org/10.1016/j.cell.2016.08.056 Find plasmids...targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids from...for targeted cytosine demethylation in mammalian cells. This plasmid is used with pcDNA3.1-MS2-Tet1-CD ...vectors and targeted delivery into hard-to-manipulate cell types. They have also fused LSD1 to x-Cas9(3.7)....modifiers may be more frequently inherited by daughter cells. For example, KRAB induced silencing was transient...enhancers. As the effects of these tools are likely cell type- and context-dependent, it may make sense to...
  24. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ... require minimal cellular perturbation. Also, this method requires live, intact cells; the sequencing ...Synthetic recombinase-based state machines in living cells” (1), into a more straight-forward, concise explanation... our paper (1), we allowed individual bacterial cells to form colonies on a plate and then we performed..., but more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The...sufficient for greater than 50 percent of RSM-containing cells in a population to correctly record each possible... were sufficient for greater than 90 percent of cells to correctly record each possible permuted substring...Synthetic recombinase-based state machines in living cells. Science. 353, aad8559 (2016). PubMed PMID: 27463678...
  25. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    ...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...biosynthesis in Drosophila melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID...
  26. Of Myc and Men

    Type
    Blog Post
    ...For about as long as the disciplines have existed, cell and molecular biologists have been continuously ... peer into the mysterious, microscopic world of cells. The development of antibodies as tools allowed ...monoclonals would help the field explore Myc’s role in cell proliferation and tumorigenesis.  Using two different...specific for human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616. https://doi.org/10.1128/mcb.5.12.3610... and immunoglobulin heavy chain binding protein. Cell 46:291–300. https://doi.org/10.1016/0092-8674(86...signal prevents secretion of luminal ER proteins. Cell 48:899–907. https://doi.org/10.1016/0092-8674(87...
  27. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...measuring protein expression or localization in cells, or even single-particle tracking of the bound complex...(1–10) and GFP(11) Versatile protein tagging in cells with split fluorescent protein. Kamiyama et al. ...tracking of protein-protein interactions in living cells. Shao et al. Commun Biol. 2021 Yujie Sun   For...bright enough for single-molecule tracking in live cells.  Controls  When designing experiments, it’s important... fragments themselves can sometimes interact in cells and produce background signal (especially for some...some older split-GFP constructs or certain cell environments). Also consider that a protein fusion may...) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent protein tags. ...
  28. Antibody Data Repositories and Search Engines

    Type
    Blog Post
    ... Immune cells Flow cytometry (potentially others) Data repository From the Human Cell Differentiation...repository   HuBMAP Consortium Healthy human cells Imaging (IHC, ICC, IF, etc.)  Data repository...
  29. CRISPR Cheat Sheet

    Type
    Blog Post
    ...nuclease target multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of...range. CRISPR can be difficult to deliver. Not all cells efficiently take up plasmids used to produce CRISPR...
  30. New Viral Vectors - Fall 2024

    Type
    Blog Post
    ...the fall colors are inspiring you to paint your cells in equally vivid colors, you'll want to check out...AAV9-X1.1 enables expression in brain endothelial cells via an injection into the systemic circulation. ...levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also...recombinase or a fluorescent marker in the various cell types of the basal ganglia. They work via newly-...
  31. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...luciferase assays require cell lysis as the most efficient means to disrupt the cell membrane and deliver ...produced. A similar approach can be used to analyze cell signaling pathways or miRNA knockdown. BRET for ...advantageous properties. This approach can improve live-cell imaging by avoiding autofluorescence and phototoxicity...can permit measurements of luminescence from live cells. Luciferase variants with incompatible substrates.../phototoxicity Not susceptible Susceptible Subcellular imaging Improving Well-established High-throughput...
  32. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...editor in mammalian cells Not all Cas13d proteins are functional in mammalian cells, but Konermann et al...make it possible to target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding... 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:...
  33. Antibodies 101: Polyclonal Antibodies

    Type
    Blog Post
    ...system, antibodies are produced by B cells. Each individual B cell produces antibodies that all recognize...also all the same isotype. But together, all the B cell clones in the immune system make different isotypes...antibodies starts with selecting just one of these B cell clones for further antibody production, polyclonal... blood from the immunized animal. The red blood cells and the serum are separated, and the serum - which...
  34. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured. Archon1 ... it. Using a robotic picking approach to select cells with variant characteristics they wanted, the team...approach to control protein-protein interactions in cells because the protein binders can be customized for...TOM20 (pTriEX Tom20 mVenus BoPD). In dark adapted cells expressing these plasmids, tgRFP fluorescence localizes...offers a way to spatially and temporally control cellular processes in vivo. Many photoactive proteins exist...photo-reversible protein-protein interactions can control subcellular localization processes in vivo. They deposited...
  35. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...Drosophila cell line, I’ve studied a transcription factor important for T cell differentiation (a cell type ...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. ...
  36. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ... can rupture E. coli spheroplasts (cells which have had their cell walls removed with lysozyme). This ...protein fusions allowed us to image R bodies inside cells, and these constructs may be useful to future scientists...applications. For example, you could use them to lyse cells (and surrounding membranes) in keeping with their...assemble and function, we might be able to endow cells with new ways to interact mechanically with their...penetrate other barriers (like vasculature or extracellular matrices) in response to pH changes. Beyond...
  37. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;156(5):935-49. doi: 10.1016/j.cell.2014.02.001. Epub 2014 Feb...Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct...screening for genes that, when turned on, make melanoma cells resistant to treatment with a BRAF inhibitor. (The...Weissman and his colleagues demonstrated in a paper in Cell that they could typically achieve 90-99% knockdown...straightforward to apply the new library for use in various cell types and in live mice. Therapeutic uses in humans...Addgene, enabling systematic investigation of the cellular consequences of repressing or inducing individual...
  38. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ... systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016/j.cell.2023.03.034 Resources...Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016/J.CELL.2013.07.018 Anzalone, A....epigenetic manipulation of prime editing. Cell. https://doi.org/10.1016/j.cell.2024.03.020 Mathis, N., Allam, A...suited for prediction of very diverse edit types and cell contexts. Figure 1: High-throughput screening...double-strand break repair pathway balance. Molecular Cell, 81(10), 2216-2230.e10. https://doi.org/10.1016/...
  39. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...Proteins in Living Cells Using Bimolecular Fluorescence Complementation. Molecular Cell 9:789–798. https...manner. This is straightforward in easy-to-transfect cell lines where plasmid dosage can be tightly controlled...fluorescence-based method for directly visualizing PPIs in cells. The ease with which BiFC can be visualized makes...visualization of protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038...(BiFC) Analysis of Protein Interactions in Live Cells. Cold Spring Harb Protoc 2013:pdb.prot076497. https...directly visualize PPIs that regulate numerous cellular functions, including ubiquitinoylation, kinase...BiFC studies used the method to interrogate the subcellular localization of interactions between basic leucine...
  40. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...ability to interact with protein partners within the cell. However, antibodies can have low affinity for these... generally bright, stable and well tolerated by cells. They can be used for protein localization, isolation...trigger protein aggregation which is toxic for the cells and affects the labeling of small structures like...team, an ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with...example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays a critical role in neural...Advances in fluorescent protein technology." Journal of cell science 120.24 (2007): 4247-4260. PubMed PMID: 18057027...of neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in the...
  41. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ..., peptides, amino acids, drugs, metal ions, and cells (even pathogenic bacteria). That range is possible... combination of SELEX and fluorescence-activated cell sorting (FACS) of E. coli transformed with aptamer...and FACS, Broccoli is inherently more stable in cells. The new aptamer also has a higher melting temperature...cassettes, thereby extending their use into mammalian cells. They’ve also dimerized Broccoli to further enhance...Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. USA 112(10) (2015): 3002...Aptamers as a Tool to Study RNA-Modifying Enzymes.” Cell Chem Biol. 2016 Mar 17;23(3):415-25. PubMed PMID... over half a century; the Addgene blog has an excellent primer and tips from experts on FRET techniques...
  42. Fluorescent Proteins: FRET

    Type
    Collection
    ...influenced by many factors, such as the organism or cell type, relative position and orientation of the fluorophores... (Unpublished) FRET Indicators - Live and Fixed Cells Additional Resources at Addgene Learn more on the...
  43. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...more about. Our post on “Making Your Own Competent Cells” has been extremely popular, generating many additional... We’re happy to see scientists making their own cells to save money and create the tools that work best...Save Time and Money by Making Your Own Competent Cells By Melina Fan Plasmids 101: The Promoter Region...
  44. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

    Type
    Blog Post
    ...Multiplex genome engineering demonstrated in human cells They used the Golden Gate cloning method to ligate...successfully to target multiple sites in cultured human cells. While it has not yet been demonstrated, they say...Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. Sakuma...
  45. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...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, ...
  46. Evolution of Lab Techniques

    Type
    Blog Post
    ...Neuroscience at Harvard University. Just as computers, cell phones, and cars become more technologically advanced...gene mutation is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with... of the mutated β globin gene that causes sickle cell anemia. The mutation happens to destroy a sequence...conducted on a sample from an individual without sickle cell anemia, two DNA fragments are observed because the... site where MstII cuts. In a patient with sickle cell anemia the enzyme does not cut the DNA so only one...Wai Kan. "A sensitive new prenatal test for sickle-cell anemia." New England Journal of Medicine 307.1 (...transgenic constructs and mice." Current Protocols in Cell Biology (2009): 19-10. PubMed PMID: 19283728. PubMed...
  47. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...pulses to the subject/cells expressing the optogenetic tool causing a change in cellular activity or gene ...positively charged ions to rapidly pass through the cell membrane and initiate an action potential - a very...such as Na+, K+, and Ca++ to passively enter the cell with the concentration gradient to depolarize it...the concentration gradient and hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine...different wavelength, turns it off, allowing the cell to be excited over a sustained period of time (Gong...affect how rapidly you can activate or inhibit a cell in succession and maintain fidelity with the stimulus...many downstream signaling mechanisms that are often cell-type specific. If it is something you’re considering...
  48. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...that it appears “newly synthesized” to the cell. Thus, the cell repairs the DNA using the U-containing strand..., which may lead to cell death rather than NHEJ repair, thus removing these cells from the edited population...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual base...relatively low frequency, especially in nondividing cells. Scientists everywhere wistfully imagined being ...editing efficiency, Komor and team wanted to force the cell to use the deaminated DNA strand as a template. ...frequency to above 30% for a variety of targets in human cell lines, with an average indel frequency of only 1.1%...modifications. CRISPR base editing persists through multiple cell divisions, indicating that this method produces ...
  49. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...complex cellular behaviors requires making complex manipulations to cells. For example: in a cell there ...TU, or even an individual part, will behave in a cell. For unpredictable cases, MoClo provides two answers...allow you to study natural genetic circuits in the cell. Natural and synthetic product production A major...biofuels, drug precursors, etc., microbially or in cell-free systems. These pathways typically involve multiple... yield a cellular phenotype, translational output, etc., which can support a larger cellular engineering...
  50. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...Deletions in Human Embryonic Stem Cells Using Type I CRISPR-Cas. Molecular Cell, 74(5): 936-950.e5 (2019). 10.1016...a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016/j.cell.2015.09.038 Harrington, B., ... Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral RNA...cleavage and is significantly less toxic to mammalian cells than Cas13 (likely due to the lack of collateral...
Showing: 401 - 450 of 813 results