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We narrowed to 819 results for: cell

Showing: 361 - 390 of 819 results

  1. NETRF

    Type
    Collection
    ... Modeling Neuroendocrine Tumors Using Adult Stem Cell-Derived Organoids Matthew Meyerson Dana-Farber Cancer...Institute Epigenetic Regulators of Intestinal Endocrine Cells and Carcinoid Tumors Qiao Zhou Harvard University...University Epigenetic Regulators of Intestinal Endocrine Cells and Carcinoid Tumors Bradley Bernstein Broad Institute...

  2. Depositor Collections

    Type
    Collection
    ...Collection Allen Institute for Cell Science Plasmid Collection New England BioLabs Cell-Imaging Plasmid Collection...Botman-Teusink Yeast FP Collection Protein Expression Cell Migration Consortium Malate Dehydrogenase CUREs ...

  3. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...holes in the cell membranes of your sample to allow antibodies better access to intracellular targets. If...allowing antibodies into the cell where they could bind to intracellular epitopes. C) Permeabilizing with...method for visualizing molecules of interest within a cell or tissue. In that introduction, we broke down the... large proteins that need a little help crossing cell membranes as they are not able to diffuse across...target protein that is still on the inside of the cell. However, assuming you do need to permeabilize, ...For example, saponin interacts with cholesterol in cell membranes and leaves many membrane-associated proteins... organic solvents. However, detergents can lyse cells or cause you to lose soluble proteins from the sample...

  4. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. PubMed...RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227.   Potential ...particularly helpful for targeting non-dividing cells, which are difficult to modify through homology-... Cpf1 orthologs known to be active in mammalian cells, LbCpf1 and AsCpf1. In both reports, on target editing... (RNPs) failed to induce off-target editing in a cell culture model. What we still don't know about Cpf1...reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016 Jun 6. PubMed PMID: 27272384...specificities of CRISPR-Cas Cpf1 nucleases in human cells." Nature Biotechnology (2016). PubMed PMID: 27347757...

  5. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

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    Blog Post
    ...metabolite in a cell. The only way to do that at scale right now is to grow up a bunch of cells, sacrifice ...capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks Foundation...as an operating system for engineering an entire cell. Typically when people use plasmids, they are trying... you could imagine engineering the entirety of a cell. That's a big long-term technical challenge. We ...That's central to everything that goes on inside a cell. Until we get a handle on precise, forward engineering...scale, it's going to be tricky to tame the rest of a cell. The BIOFAB plasmids represent the first systematic...transistor-like digital genetic switches that enable cellular computing. These logic gates built from transcriptors...

  6. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ...genetic cargo, typically an expression construct, to cells/organisms. This is often cDNA, shRNA, or something...perform very similar functions to infiltrate the host cell and deposit viral cargo. Gene expression While AAV...what will be delivered and expressed in the host cell from the transfer plasmid. Viral packaging and entry...altered tropism of AAV to target different tissue or cell types. AAV also requires some back up from adenovirus...promoter that you wish to deliver to your target cells. In adenovirus, the ITRs serve as a type of origin...deficient. This is typically supplied by the production cell line (HEK-293) during production. Fun fact: There...

  7. 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...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...

  8. Screening for Successful Genome Editing with Digital PCR

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    Blog Post
    ...to quantify rates of mutation in a bulk or single cell-derived genomic DNA sample as part of a precision... successfully analyzed gDNA from a few thousand cells from single wells of a 96-well plate without prior... that they are easily able to distinguish single-cell derived samples with mono-allelic mutations versus...Isolation of single-base genome-edited human iPS cells without antibiotic selection. Nat Meth 11, 291–293... and Highly Specific Screening of Genome Edited Cells. PLoS ONE 11, e0153901–17 (2016). PubMed PMID: 27089539...mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology 31, 822–826 (2013). PubMed...

  9. Plasmids 101: Cre-lox

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    Blog Post
    ...transgenic animals, embryonic stem cells, and/or tissue-specific cell types: Cre-lox recombination. What...downstream of promoters that are active only in certain cell or tissue types, during certain stages of development... recombinase can be expressed only in specified cells or at specified times. Combining this with some ...has been used to successfully introduce Cre into cells of interest. What if I need two separate recombination...2. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Sauer...

  10. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...continuously synthesized and degraded within the cell, so at any given time, an FP-chimeric protein may...Since emission of green light is more phototoxic to cells, most scientists working on in vivo imaging models... this is not ideal for experiments in mammalian cells. Dendra2 is another monomeric PA-FP which unlike...tetrameric. Tetrameric PA-FPS are better for whole cell imaging and studying organelle trafficking. Monomeric...the photoactivation step can actually harm your cells before your experiment even begins! If you are performing...optical live imaging of proteins, organelles, and cells. Similar to FPs, PA-FPs allow for non-invasive labelling...dynamics of mitochondria in healthy and apoptotic cells (9). PA-FPs have helped to revolutionize the field...

  11. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...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...

  12. Choosing Your Perfect Empty Backbone

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    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 (...

  13. What's New in CRISPR - Spring 2019

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    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...

  14. Hot Plasmids: Winter 2025

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    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...

  15. MXS Chaining

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    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...

  16. 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...

  17. 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...

  18. Viral Vectors 101: Virus Safety

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    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...

  19. 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...

  20. Cancer Research Plasmids and Resources

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    Collection
    ...important to normal cell signaling and physiology, and, when dysregulated, can enable cancer cell survival. Although...that is essential for regulating the growth of the cell. p53 p53, the most frequently mutated gene in human...

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

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    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...

  22. Visualizing Translation at the Single Molecule Level

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    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...

  23. 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...

  24. CRISPR Plasmids - Bacteria

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    Collection
    ...to activate programmed cell death or dormancy for phage-infected bacterial cells so as to limit the spread...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ...Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both...

  25. Plasmids 101: Using Transposons in the Lab

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    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...

  26. CRISPR Plasmids - RNA Targeting

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    Collection
    ...to activate programmed cell death or dormancy for phage-infected bacterial cells so as to limit the spread...population. Type VI enzymes that function in mammalian cells can be used to attentuate RNA levels. In mammalian...

  27. Plasmids 101: Modular Cloning Applications and Kits

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    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...

  28. Educational Planarian Tools for Teaching Developmental Biology Techniques

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    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...

  29. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

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    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...

  30. 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...
Showing: 361 - 390 of 819 results