We narrowed to 888 results for: URE

...that overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule...specific place and time. Future studies with these CIB1-Rabs in other contexts are sure to reveal even more...cytoskeleton. They provide cells with physical structure, separate chromosomes during mitosis, support ...regions, laser ablation can damage other cellular structures, and genetic mutations are not easily reversible...roles played by microtubules, Opto-katanin will surely be useful for a wide variety of experiments. Opto-RhoGEFs... GEF to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized ...your cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity....

...variants:   Figure 2: Inner mitochondrial membranes imaged with structured illumination microscopy...incubation, even with multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, ...primary cells using peptide-assisted genome editing. Nature Biotechnology, 10.1038/s41587-023-01756-1. https...Although it quickly saw wide interest, its dimeric nature can interfere with some applications. Several labs...molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038/s41592-023...2023). A monomeric StayGold fluorescent protein. Nature Biotechnology, 1–4. https://doi.org/10.1038/s41587...observed in young macaques and vascular bias in mature marmosets. CAP-Mac also robustly expressed fluorescent...

...gleaned from the literature and engagement with researchers who study the protein or structure of interest....alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope represents...case of protein tagging to elucidate cellular structures. Overexpression methods flood the cell with protein...visualize the precise location of the protein or structure of interest. Endogenous gene tagging is an ideal.... Furthermore, the quality control necessary to ensure the edited cells are behaving normally can be prohibitively...tagged are widely recognized markers of cellular structures, we provide guidance on how to make your own ... Collection Fluorescent tagging strategy Figure 1: Gene editing is the addition of a useful coding...

...the proper controls needed to pressure-test your FRET assay to make sure it is working? This article addresses...resonance energy transfer measurements of ryanodine receptor type 1 structure using a novel site-specific...donor/acceptor fluorophores (Stryer, 1978), a measurement that is unaffected by intervening proteins, lipid...blog has an excellent primer on FRET and how to measure it). Molecular mailbox: How to get your fluorophore... on the same street as the donor), then FRET measurements are possible. In fact, even if the majority ...properly targeted donor/acceptor pairs, then any measureable FRET will be swamped out by a sea of uncoupled..., but even these modifications can affect the structure or function of certain proteins. Rather than shove...

...introduced mutations that drive leukemia (Figure 3). Figure 3: Using LARRYv2 libraries for state-...expansion cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures from days...maintaining high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments ...yellow fluorescent proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038... an RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral...less than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...

...commonly called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of AAV production...understand how different titers are measured. A TCID50 measurement reports an infectious titer, meaning...Whitney, O., Walther, N., & Maurer, A. C. (2023). Transgene Expression in Cultured Cells Using Unpurified ...with standard molecular biology tools and tissue culture experience. Here, we’ll go over the basics of rAAV...rAAV production.  Of course, you will want to make sure that your final stocks only contain rAAV. To minimize...contamination, use appropriate decontamination procedures (Korte et al., 2021) for all disposable items...into contact with the rAAV and apply common cell culture practices diligently throughout the production ...

... efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https... evaluate all of the pegRNAs in one experiment (Figure 1). Through this method, we found key determinants...Once we had our datasets with all the different features and editing efficiencies, we could train our prediction...very diverse edit types and cell contexts. Figure 1: High-throughput screening of pegRNA efficiency...downstream flanking sequences to their input string (Figure 2). This string can then be used as input for the...e.g., top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with...Mathis et al., 2024) and concurrent research by the Shendure lab (Li et al., 2024) have highlighted how chromatin...

... SDS-PAGE, measure the protein content and normalize to ensure equivalent loading. Denature the samples...antibodies.   Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated...denaturing western, the protein is denatured to its primary structure and separated by size with smaller...western, the protein’s secondary and tertiary structures remain intact and the protein is separated through... expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack...its native state while a western blot detects denatured polypeptide chains. The protein regions available... antibody validation. This level of validation ensures that the antibody is staining the intended target...

...of Addgene’s work culture. Our developers are a diverse group of ethnicities, cultures, genders, educational... for working on a new feature. We start with a story that describes the feature and what the criteria ...stakeholder accepts the new feature as done and we’re onto the next story. Figure 1: Addgene’s Agile Approach...working on a feature, sometimes we have to take short cuts or add a band-aid to get a feature to work. We...number of stories we commit to for each sprint, to ensure we will have the capacity to finish all of the ...criteria is for accepting the feature as done correctly. These initial stories are usually a result of collaborations...step-by-step instructions on how to demonstrate the feature. For example, we recently reduced the number of...

...sequence (Zong et al., 2018) (see CBE-3 in Figure 3). Figure 3: Activity windows of plant base editors...excision repair mediated by uracil glycosylase (Figure 1). The presence of a uracil glycosylase inhibitor...generation of the desired T-A base pair. Figure 1: Transition mutation (C>T, G>A, A>G, and T>C)... that compete with the desired outcome are not pictured. Adenine base editors (ABE) What if you have...satisfactorily accepts DNA as its substrate (see figure 2).          The artificially evolved adenosine...genomic A-T base pair is converted to a G-C pair (Figure 1). As the cellular inosine excision repair is ...base editing platforms for plant systems.  Figure 2: Plant base editor platforms and their molecular...

...better at 30°C or room-temperature, which is accomplished by growing your culture to the desired density...previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli for DNA ...except ** which are K12 Strain Resistance Key Features Genotype Use BL21 (DE3)   Basic IPTG-inducible...Denotes the presence of an additional plasmid-- make sure to maintain this by growing on appropriate media...timing is important, as you typically want to make sure your cells have first reached an appropriate density...expression. If you wait too long to induce, your culture will start collecting dead cells, and, conversely...too early as there are not enough cells in the culture to make protein.  The DE3 lysogen/T7 promoter combination...

...acceptor fluorophore can be measured using microscopy techniques. FRET measurement sensitivity makes it suitable...one of the first techniques which enabled the measurement of single molecule interactions in living cells...living cells. Since then FRET has also been used to measure conformational changes, cleavage activity using...is still one of the best and most used pairs to measure FRET. The table below lists plasmids that can be...and Venus Other issues that can affect FRET measurements include: the brightness of a fluorophore pair...between FRET efficiency and cross-talk. Methods to Measure FRET for Cell Biology Studies Several methods have... have been used over the past 20 years to measure FRET and there is not one that is better than another...

...environments: corrosive pH ranges, crushing pressures, near-melting temperatures, tons of radiation, or freezing ...Sreepadmanabh M from IISER Bhopal. All illustrations / figures featured were designed by our team member Kallol Mozumdar... take bacteria out of their optimally suited temperature ranges and make them grow better at suboptimal...cold-loving) bacteria. These bacteria inhabit cold temperature locales and psychrophilic enzymes help them survive...next-door microbes which grow optimally in moderate temperatures (usually defined between a fairly nippy 20°C...materials degrade at a much slower rate under cold temperatures, and frigid conditions decrease the potential...mesophilic host systems. The so-called “regular” temperatures of growth, say around 37°C, is a far cry from...

...method does provide a quantitative measure of autophagy, it only measures bulk autophagy (as opposed to giving... Long-lived protein degradation: This approach measures the degradation rate of radio-labeled long-lived...long-lived proteins as a proxy for autophagy. Cells are cultured with isotope-labeled amino acids for several hours...by measuring the amount of radioactivity in the culture supernatant (this is indicative of protein degradation...out of this assay. This is also a quantitative measure of autophagy, but, like the protein degradation...degradation assay above, it’s slow and only measures bulk autophagy. Morphological assays Electron microscopy...method relies on the identification of autophagic structures based on morphology. Autophagosomes are relatively...

...articles, only 46% could be reproduced. In a recent Nature report from researchers at Amgen, it was stated...that only 11% of the academic research in the literature was reproducible by their groups. Finally results..., it will always be a part of everyday work to capture ideas, write down certain notes or brainstorm, ...functionalities i.e. main priorities of different ELN features, ranging from whether respondents saw them as ...term, so they need a solution for which they are sure that there will be no sharp changes in costs once...researchers to store everything in one place. From pictures to excel sheets, researchers today generate vast...Following these findings, we defined the priorities for future ELN development and developed the SciNote Electronic...

...with CRISPR–Cas13. Nature, 550(7675), 280–284. https://doi.org/10.1038/nature24049 Altae-Tran, H., Kannan... ideas for your own research! Classification Structure Principles of CRISPR classification While CRISPR...six types, 33 subtypes, and 17 variants. Figure 1: Overview of the CRISPR classification hierarchy...using a Cascade complex is tricky, as you need to ensure every unit of the complex expresses at similar ...components present in the Cascade complex. Figure 2: Schematic of a Cascade complex (blue subunits...CRISPR types do, due to a lack of canonical CRISPR features (Pinilla-Redondo et al., 2019; Moya-Beltrán et... most often found on plasmids. The many unique features of IV-A and IV-B have led to the hypotheses that...

...analyzes publications and figures using machine learning to extract important features such as influential ... top of the ever expanding body of biomedical literature! PubMed PubMed is the classic search engine ...eventually replace the existing version of PubMed.  Key features This new version has an improved search algorithm...updated. This Twitter list contains >300 Twitter literature bots, including a bot for biosensor papers, microbe...Semantic Scholar Semantic Scholar is an AI-backed literature search developed by the Allen Institute for Artificial...that are more relevant to your search terms. Key features Semantic Scholar has several ways to focus your... populated at the top of this filter. A unique feature of Semantic Scholar search results are the topic...

...noise and ensure you are measuring the events-of-interest. Sensors suitable for ER measurements include:... requiring only a simple measurement. However, single wavelength measurements are more susceptible to ... RE. Structure-and mechanism-guided design of single fluorescent protein-based biosensors. Nature chemical...subtype) and specific subcellular compartments for measurements. All of the sensors discussed below fall into...slower). GCaMPf may be suitable for very rapid measurements, like action potentials in neurons. jGCaMP7 ...additional versions are available to simultaneously measure calcium changes in the ER, cytosol and mitochondria...you combining your calcium reading with other measurements, specifically fluorescent ones? Many of the ...

...to compare exposure times of our samples. A prominent, strong signal has a shorter exposure time than ... be essential to your new research project, but unsure which one will mark your target the best, or even...The strength is based on binding specificity and exposure time. We first have a negative control which does...align with what the atlases (mentioned above) and literature suggest, providing an expectation and comparison...imaged pattern does NOT align with the atlases/literature and is unrelated to the target protein, we categorize... goldilocks antibody selection process The exposure time of the primary antibody compared to negative...control also determines the signal’s strength. Exposure time is the amount of time the sample needs to...

...using cloning rings. A picture of a 60 mm dish ready for picking is shown in figure 1.   Find Additional... Protocol Pages Agarose cloning method procedure The procedure (illustrated in the YouTube video below...well plate is also prepared with about 200 ul of culture medium containing serum in each well. A 200 ul ...both Hepes and bicarbonate) is brought to room temperature and placed in the hood. The medium from a 10 ...is capped, swirled and tested for the correct temperature by placing on the wrist or cheek. Within less...warm but not hot- and still liquid- it is rapidly poured onto the middle of the plate, where it immediately...the agarose cloning method Test your agarose temperature. Cloning cells using an agarose overlay does ...