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We narrowed to 811 results for: LARS

Showing: 261 - 280 of 811 results

  1. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    Published
    March 19, 2019, 1:31 p.m.
    ...potently inhibit type II CRISPR systems including the popular S. pyogenes Cas9 (Rauch et al., 2017, Hynes et ... on a single plasmid (Grefen and Blatt, 2012). Similar to traditional Gateway™ cloning, the GOI first ... (FRET, Heckler et al., 2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and...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...the mitochondria, because cPYP binds to BoPD. A similar set of newly identified light- and dark- binder...

  2. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    Published
    Nov. 21, 2019, 1:53 p.m.
    ...can be particularly useful in academic labs when creating shared experimental protocols. Clarify: Asking...setting, where student and postdoc turnover happens regularly and it behooves everyone in the lab to be using...Asking for clarification during a meeting can help get everyone on the same page. Others in the meeting might...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing 8-10 ...understanding you can be incredibly frustrating. Clarifying the words —> decreasing frustration —> increasing...

  3. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...: IL-36 biology in generalized pustular psoriasis Generalized pustular psoriasis is a chronic autoinflammatory...in this educational video. “SLC6A8 belongs to a large family of solute channels that are responsible for...outside of the cell and translate those signals into cellular responses. For example, GPCRs and their downstream...different receptors upon stimulation. “To study cellular transduction events, we will utilize BRET-, FRET...intensity-based signaling biosensors to monitor intracellular signaling,” says Hébert. Previous research has...

  4. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...reagents.  Affinity Reagents  Affinity reagents are molecular tools that have the ability to specifically recognize...affinity reagents is their size. Antibodies are large enough that their bulk can be a barrier at times...difficult to source, may not be readily available for a large number of targets, and can require specific conjugates...   Types of Affinity Reagents  (Listed in no particular order.) Fab fragments are created through enzymatic...variables (scFvs) are ~27kD engineered fusion proteins similar to a Fab fragment but created through engineering...eukaryotic cells, allowing for them to be used as intracellular inhibitors.  Aptamers are not proteins at all...

  5. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...encode proteins. They play important roles in many cellular and regulatory processes, but much of their functionality...removing the full ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for ... same time (Liu et al. 2018). However, removing large lncRNA fragments with multiple gRNAs poses the risk...Zibitt et al. 2021). Mutating splice sites is particularly effective as it can result in the retention ...Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (D) Knock-in: insertion of a synthetic...used to target lncRNAs with high specificity, particularly in applications requiring smaller delivery systems...

  6. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...developed third option: Fragmid. Fragmid is a modular vector assembly process designed for a quick, plug-and-play...vectors in five days. Did you say… easier? Fragmid’s modular assembly structure takes a lot of the design work... And it truly is plug-and-play. Because of its modular cloning approach, it’s easy to switch out reporters... by phiC31   The Fragmid website While the modular cloning approach is key to the Fragmid system, ..., M. A., Green, T. M., & Doench, J. G. (2024). Modular vector assembly enables rapid assessment of emerging....2024.100519 More resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden...

  7. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ... to interact by stimulation with blue light.  Similarly, light can also be used to control homo-oligomerization...distribution of clusters without accumulation in subcellular compartments such as nuclear speckles. Furthermore...OptoSTIM1), caused two-fold faster changes in intracellular Ca2+ levels than the original version containing...CRY2clust should prove a useful tool to further probe cellular function. The CRY2clust Plasmid can be found here... commonly observed with direct stimulation are largely absent in the soCoChR-expressing system. Taken ...interactions. We’re excited to see how you adapt similar FLINC biosensors to work in your lab. Find the ...

  8. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...and TadA7.10, they designed one set of ABE8’s similarly (ABE8.x-d). In order to assess the effect of removing...you need to modify multiple genes to generate a cellular phenotype. Again ABE8.20-m was the winner. It ...editing in ABE8s While base editors are precise molecular tools, they do have undesired off-target editing...have been described to introduce base changes in cellular DNA and RNA dependent or independent of the gRNA..., and a thoughtful selection of guideRNAs. Of particular note, ABE8s reported in this work did not cause...sites that used to be difficult to target. In particular, their ability for more robust editing in human...

  9. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ...genes in particular tissues, but they are costly and time-consuming to generate, especially for large animals...Adeno-associated virus (AAV) is a popular tool for gene delivery, but it has a drawback: how do you ensure... it to? Knowing that a gene is expressed in a particular cell type is important not only for translational...types.  Botond Roska’s lab at the Institute of Molecular and Clinical Ophthalmology Basel recently took...Besides restricting AAV transgene expression to particular cell types or classes, these synthetic promoters...

  10. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    Published
    Aug. 9, 2022, 1:15 p.m.
    ... very specific biological targets. However, as popular and useful antibodies are, they are in a midst ...as non-human primates and humans. Since we need larger-volume antibodies for thicker tissues and whole... immunoglobin component that can be produced in large “copies.” This homogeneity circumvents the batch-to-batch...reproducibility and cost-effectiveness for broad and large quantity uses. We therefore use both rAb and mAB...same target to see if the results are the same/similar, emphasizing the antibody’s specificity. Strength...from the background noise. If the sample images similarly to the negative control, that informs us that ...

  11. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...polyadenylation signal by using in vitro analyses. Molecular and Cellular Biology. 1992. PMID: 1333042. PubMed Central... thought that poly(A)s are used to control the cellular concentration of regulatory RNAs and may additionally...secondary structure to induce transcript cleavage, similar to the Rho-independent termination found in prokaryotes...terminate transcription. Polymerase I uses a process similar to the prokaryotic Rho-dependent mechanism, whereas...Terminators: The RNA 3′-End Chronicles. Journal of molecular biology. 2011. PMID: 21439297. PubMed Central ...

  12. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ...Simon! Derek P. Simon received his PhD in Cellular and Molecular Biology from the University of Michigan...blogger Derek Simon. The surgeries and standard molecular neuroscience validation experiments we discussed...cemented to the animal’s skull using dental cement (a similar procedure as implanting a guide cannula). A fiber... Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje cells. The Journal of biological chemistry...PMCID: PMC3466483. 4. Kim TI, et al. Injectable, cellular-scale optoelectronics with applications for wireless...

  13. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    Published
    Nov. 5, 2024, 8:32 p.m.
    ...cells are a great option for viral production, particularly for labs that already do mammalian cell culture...for the generation of viruses, though they are a popular choice for protein production. The most commonly...infect insects. This is what makes insect cells a popular choice for protein production. AAV and lentiviruses...systems. Baculovirus transfer plasmids have a much larger packaging capacity — up to 38 kb! Generally, insect...in between passages to eliminate dead cells and cellular debris. While adding time (about three to four...supernatant in a centrifuge to remove any unwanted cellular debris, as you would when collecting a lentivirus...

  14. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems, these...nucleotide sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes...targeting constructs. In the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the ... the cells that have randomly integrated all or large portions of the plasmid. Deliver your construct ...insert GFP into the genome, we’d create a plasmid similar to the one shown below, where the sequence of GFP...CRISPR is very good at making knockouts, knocking in large sections of DNA can be more difficult. Addgene depositors...

  15. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ... that can bind effectively to protein targets, similar to how antibodies do. The structure of the common...(~150 kDa) and Y-shaped because it contains two large heavy chains and two small light chains that stick...antibodies to get to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure...optimally binding sybody, you want to start out with a large and diverse pool of sybodies that hypothetically...because it has the advantage of starting out with a larger initial starting library size (up to 1012 members...Characterization of Enzyme Variants. In: Methods in Molecular Biology. Humana Press, pp 133–148. https://doi.org...

  16. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...already a packaging capacity larger than AAV vectors. This capacity is large enough to carry the Cas9 gene...compared to other viral vectors, AdVs can provide a large transgene packaging capacity. For instance, with...non-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding TALENs... B-thalassemia, Sickle Cells Disease, Duchenne Muscular Dystrophy, Alpha1-antitrypsin deficiency (most... animals in order to create animal models for particular cancer. That’s exactly what the lab of Andrea...

  17. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ...into embryos and then become larvae. There are three larval stages where larvae molt in order to continue...continue growing in the next larval stage. After the third larval stage, Drosophila will pupate, which takes...fertilized egg by its mother. Inside the pupal case, the larva will metamorphose into its adult form. In all, it...

  18. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ... might need to use multiple techniques for a particularly stubborn prep. Check the bacterial strain Increase...strains, including HB101 and derivatives, release large amounts of carbohydrates that can inhibit the cell...levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression...choice of strain will ultimately depend on the particular features of your plasmid, it is advisable to ...coli strains. Increase the volume Starting with a larger culture volume can improve plasmid yield. Our lab...gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around 220 RPM,...

  19. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ...depositing lab. There may be other plasmids with similar names in the collection, but you can use this name...some of Addgene’s plasmids is an indication of popularity. A yellow flame indicates that a plasmid has ... on our website, one of the available filters popularity (e.g. only showing those plasmids which have ... than 100 times).     I guess it's cool to be popular, but how do I know if a plasmid is actually useful...If the depositor has indicated that there is a particular mutation found within the gene/insert, this information...means you can see which labs have already used a particular plasmid, and which experimental systems it was...

  20. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...role of individual genes or entire pathways in a particular disease process. While CRISPR is certainly not... mutant cells to identify genes involved in a particular phenotype. CRISPR can be readily scaled up for...fluorescence-based cell sorting and screened for a particular phenotype. For example, mutant cells can be used...cell types that are of low abundance are not particularly well suited for genome-wide screening. Avoid...targeting the same gene can be strong evidence that a particular gene is actually important for a given phenotype...Lentiviral CRISPR Libraries. Current Protocols in Molecular Biology 115: . https://doi.org/10.1002/cpmb.8 ...
Showing: 261 - 280 of 811 results